Reducing oxidative stress by κ-carrageenan and C60HyFn: The post-thaw quality and antioxidant status of Azari water buffalo bull semen.

Azeri water buffalo is a species of great interest due to the high quality of its products such as milk. Due to the decreasing trend of its number and risk of extinction in the future, our attention is directed towards ensuring the preservation of its genetic reserves by keeping its sperm. Using antioxidants in semen extender is one of the ways to reduce the detrimental effects of freezing process on post-thawed quality of spermatozoa. This study was conducted to determine the effect of κ-carrageenan (k-CRG) and C60HyFn supplemented semen extender on the quality of post-thawed Azari water buffalo spermatozoa. A total of 30 semen samples were obtained from three buffaloes using an artificial vagina (twice a week for five weeks = 10 replicates). The samples (n = 3) from each replicate were pooled and divided into equal aliquots to prepare 14 extender groups, including control (C), k-0.2, K-0.4, K-0.6, K-0.8 (containing 0.2, 0.4, 0.8 mg K-CRG/mL, respectively), C-0.1, C-0.2, C-0.4, C-0.8, C-1, C-5, C-10, C-20, and C-40 (containing 0.1, 0.2, 0.4, 0.6, 0.8, 1, 5, 10, 20, 40 µM C60HyFn, respectively), and then frozen. After thawing, motility and velocity parameters, plasma membrane integrity (PMI) and functionality (PMF), DNA damage, Hypo-osmotic swelling (HOS) test, malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase glutathione activities and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging were evaluated. In vivo fertility was compared between k-0.6, C-1 and control groups. 60 buffalo were inseminated 24 h after the onset of estrus. The diagnosis of pregnancy was performed rectally at least 60 days after fertilization. Total and progressive motility and velocity parameters were improved by k-0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups) compared to the other groups. Plasma membranes integrity and PMF were improved by k-0.4, k-0.6, C-0.4, C-0.8, C-1, C-5, and C-10 groups compared to other groups, while in terms of sperm DNA damage K-0.4, K-0.6, K-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups showed better results compared to the control group. The evidence also showed that k- 0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve TAC, and decrease MDA levels. Also, k-0.4, k-0.6, k-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve GPx, CAT, and GSH levels, but no significant difference was found regarding SOD compared to the other groups. DPPH scavengers were tested by K-0.6, K-0.8 and C-1, C-5, C-10, C-0.8, C-0.4 and C-0.2 groups and compared to other groups improved. The fertility rate [70% (14/20)] was higher in C-1 than other groups. To conclude that k-CRG and C60HyFn supplementation can increase the quality parameters of cryopreserved buffalo semen after thawing and that 1 M C60HyFn can increase in vivo fertility of buffalo semen.

Changes in the capacity of the vagina and the compliance of the vaginal wall in ovariectomized, normal cyclical and pregnant ewes, before and after treatment with exogenous oestradiol and progesterone.

The 'capacity' of the vagina and the compliance of the vaginal wall was measured in bilaterally ovariectomized ewes (n = 7) before and after treatment with exogenous oestradiol and progesterone; in nulliparous ewes (n = 7) at oestrus (Day 0) and dioestrus (Day 10) and during pregnancy, and in another group of pregnant ewes (n = 15) treated with exogenous oestradiol and progesterone. Measurements were remarkably consistent within individual animals but there were considerable differences between individual animals. The 'vaginal capacity' and the compliance of the vaginal wall were greater at oestrus than during dioestrus. In the same seven ewes, which were studied during their first and second pregnancies, the 'capacity' of the vagina increased whereas the compliance of the vaginal wall declined; from 90 days to term both parameters remained fairly constant. For the first 2 months of gestation the vaginal capacity was greater in year 2 than year 1 but this was reversed during the last 3 months. The compliance of the vaginal wall was significantly greater (P < 0.0001) in year 2 than year 1 at all stages of pregnancy. In ovariectomized ewes, progesterone only significantly increased the vaginal capacity at the highest dose rate (viz. 100 mg); the compliance of the wall was reduced at the 25 and 50 mg dose rates. Oestradiol produced an inconsistent dose response effect; whilst 5 mg and 20 mg had no effect upon the vaginal capacity, the 10 mg dose rate significantly reduced it. Similarly, the highest and lowest dose rates reduced the compliance of the vaginal wall but the 10 mg dose rate increased it. At 90 and 120 days of gestation, both 5 mg oestradiol and 100 mg progesterone increased the vaginal capacity but reduced the compliance.