Methanolic pomegranate dried peel extract improves cryopreserved semen quality and antioxidant capacity of rams

Objective:To select the appropriate concentrations of methanolic pomegranate extract supplemented in rams’semen extender for obtaining the best-cryopreserved semen quality.Methods:Tris-based semen extender was supplemented with 0.0,0.40,0.48,and 0.56 mg/mL pomegranate peel methanolic extract to extend semen collected from five native rams twice weekly for two months(n=80).Pooled(n=16)post-thaw semen characteristics were determined.Thawed seminal plasma of all supplemented and control groups were used to measure malondialdehyde(MDA),nitric oxide(NO),glutathione peroxidase(GPx),superoxide dismutase(SOD),ascorbic acid,zinc,copper,total cholesterol,low-density lipoproteins(LDL),lactate dehydrogenase(LDH),and alkaline phosphatase(ALP).Results:The supplementation of Tris-based semen extender with 0.48 mg/mL semen extender resulted in the highest post-thaw sperm total motility(P<0.001),sperm progressive motility(P<0.001),live sperm(P<0.001),sperm plasma membrane integrity(P<0.001),acrosome integrity(P<0.001),SOD(P<0.05),zinc(P<0.001),total cholesterol(P<0.001),and LDL(P<0.001)with the lowest percentage of abnormal sperm morphology(P<0.001),the lowest lipid peroxidation(MDA,P<0.01),ascorbic acid(P>0.05),and LDH(P>0.05).Conclusions:Pomegranate peel methanolic extract 0.48 mg/mL supplemented to Tris-based semen extender of rams is the best enrichment in preserving the sperm post-thaw characteristics via improving biochemical profiles and antioxidant capacity.

Testicular vascularization at two locations in relation to hormonal levels,and pixel echotexture in bulls at different ages

Objective:To assess the hemodynamic changes that occurred in the main testicular artery at two different locations such as supra and marginal ones in bulls at three different ages.Methods:Eighteen Baladi bulls were divided according to their age into three different age groups:1 year[n=6;groupⅠwith bodyweight(380±10)kg],3 years[n=6;groupⅡwith bodyweight(570±10)kg],and 6 years old[n=6;groupⅢwith bodyweight(650±10)kg]Baladi bulls.Circumference of the scrotum,thickness of the mediastinum,testicular dimensions,vascularity,and hormonal levels were measured.Testicular hemodynamics were assessed by Doppler ultrasound scanning.Results:Testicular width,length,and volume were significantly different among the three age groups,with the highest in bulls of groupⅢ(P<0.05).The bulls in groupⅢhad significantly higher mediastinal line thickness and scrotal circumference than groupⅠ(P<0.05),but there was no significant difference between groupⅡand groupⅢ.Bulls in groupⅡshowed a decline in Doppler indices(resistance and pulsatility indexes)at both testicular branches compared to the other two groups(P<0.05).Supra and marginal end diastolic velocities in the three age groups were significantly different,with the highest in groupⅡ(P<0.05).Time average velocity(cm/s)was significantly elevated in groupⅡcompared to the other two groups(P<0.05).Both testicular echotextures were significantly decreased in groupⅡ(P<0.05).Estradiol and nitric oxide metabolites were significantly high in groupⅡcompared to the other two groups(P<0.05).Meanwhile,the level of plasma testosterone was maximum in groupⅢ(P<0.05).Positive correlations were found between supratesticular artery resistance and pulsatility indexes(r=0.81;P<0.001),while both resistance and pulsatility indexes had correlated negative with estradiol(r=-0.71 and r=-0.91;P<0.001),and nitric oxide metabolites(r=-0.92 and r=-0.72;P<0.001).Conclusions:Three-year old Baladi bull has the lesser Doppler indices with lesser echotexture and greater nitric oxide with estradiol concentrations,which directs a significant ability for the thermoregulation process.These parameters will help in breeding selection.

INRA82 extender enhances semen quality in ram under cooled and cryopreserved stages

Objective:To investigate the effect of INRA82 extender on ram semen quality preserved in cooled and frozen conditions in comparison with Tris-based extenders and to select the suitable extender for ram semen preservation.Methods:Semen samples were collected and pooled from Awassi rams(n=5)and divided into three aliquots.Each aliquot was diluted with one of the following extenders:(1)Tris-citrate glucose,(2)Triscitrate fructose,and (3)INRA-82.For liquid storage,diluted samples were kept at 5℃ for 72 h.Progressive motility was measured at 0,24,48 and 72 h after cooling.Besides,viability,morphology,membrane,acrosome and DNA integrities were evaluated at 24 h post-cooling.For cryopreservation,diluted samples were cooled,equilibrated,loaded and frozen in liquid nitrogen.Thawed samples were evaluated in the same manner as cooled conditions.Results:Seminal characteristics were improved after cooling and thawing in samples diluted with INRA-82 compared to those diluted with Tris-citrate glucose or Tris-citrate fructose(P<0.05).Conclusions:Dilution of ram semen with INRA-82 improves semen parameters.Hence,INRA-82,as a stimulating diluent,can be successfully used to reserve viability and sustainability of cooled and cryopreserved ram semen.

Hemodynamic changes in arterial flow velocities throughout the first six months of pregnancy in buffalo heifers by Doppler ultrasonography

Objective: To assess blood flow parameters as well as Doppler indices at the first six months of pregnancy in buffalo heifers. Methods: A total of 15 healthy, cycling, buffalo heifers were examined twice per month. Examination of Doppler ultrasonography started from the first month till the sixth months of pregnancy. All animals were subjected to transrectal Doppler ultrasonography to assess ovarian and uterine blood flow. Resistance index, pulsatility index, peak systolic velocity, end diastolic velocity, and blood flow rate were measured of both ipsilateral and contralateral to the fetus side. Results: The resistance index was positively correlated with the pulsatility index (r=0.62, P<0.01) but negatively correlated with all other parameters. For both ovarian and uterine arteries ipsilateral and contralateral to the fetus, there was a reduction in the resistance index, with a significant increase (P<0.01) in peak systolic and end diastolic velocities in both ovarian and uterine arteries. The peak systolic and end diastolic velocities of the ipsilateral ovarian and uterine arteries increased linearly till the six months of pregnancy with increased growth demands of the fetus. The same for the uterine branch of the ovarian artery was done in the middle uterine artery and umbilical artery ipsilateral to the fetus side. Conclusions: This study provides reference data of the hemodynamic changes in both ovarian and uterine arteries that could be a valuable tool to evaluate all hemodynamic changes in the developing placenta/fetus. Transrectal Doppler ultrasonography proves to be a useful non-invasive method to assess utero-ovarian blood flow during pregnancy.