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20222
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Subject: Veterinary Physiology × End date: 2022 × Start date: 2020 × Type: Thesis × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    Effect of selenium and zinc-oxide nanoparticles on cryopreserved semen quality and fertility of Assam hill goat
    (College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2022) Abedin, Sayed Nabil; Baruah, Anubha
    Nanoparticles (NPs), due to their smaller size and unique surface properties can be incorporated into a variety of reproductive biology procedures. The present investigation was carried out from September, 2021 to July, 2022 on four (4) Assam Hill Goat bucks (10 ejaculates per buck) to investigate the effect of supplementing zinc oxide (ZnO) and selenium (Se) NPs in TRIS extender on seminal attributes, lipid peroxidation (LPO) profile, antioxidant enzyme activities viz., superoxide dismutase (SOD), catalase (CAT) and Glutathione-S-transferase (GST), relative heat shock protein (HSP) mRNA levels and fertility of cryopreserved Assam Hill Goat semen. The size morphology and zeta potential values of ZnO and Se NPs were evaluated. Qualified semen samples were divided into five (5) aliquots and then diluted in TRIS extender containing ZnO and Se NP supplementation at different concentrations (T0: control; T1: 0.1mg/mL ZnO NPs; T2: 0.5 mg/mL ZnO NPs; T3: 0.5 μg/mL Se NPs and T4: 1 μg/mL Se NPs). Diluted semen was packed in 0.25 mL straws and then stored in liquid nitrogen. After thawing, post-thaw attributes viz., motility, viability, morphology, plasma membrane integrity (PMI), DNA integrity and mitochondrial membrane potential (MMP) were evaluated. The different treatment groups were also checked for potential NP internalization under transmission electron microscope (TEM). Lastly, straws from the best among the ZnO and Se NP treatments were used for artificial insemination (AI) in does (n=35) synchronized by Ovsynch protocol. Results showed that ZnO and Se NPs were poly-crystalline in nature with particle size below 100 nanometers. The evaluated post-thaw sperm in vitro attributes were significantly (p<0.05) higher in groups containing ZnO and Se NPs supplementation in comparison to control group. Overall, ZnO NPs @ 0.1 mg/mL (T1) had significantly (p<0.05) higher post-thaw sperm in vitro attributes in comparison to Se NPs @ 1 μg/mL. ZnO and Se NP supplementation also significantly (p<0.01) lowered cryocapacitated (B and AR pattern) spermatozoa in comparison to control. The antioxidant enzyme activities (SOD, CAT and GST) were significantly (p<0.001) higher in T1 in comparison to T0. The LPO was significantly (p<0.001) lowered in T1, T2, T3 and T4 in comparison to T0. The leakages of functional enzymes viz., aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were significantly (p<0.001) lower in T1 in comparison to other groups. Post-thaw sperm motility and MMP had a highly significant(r=0.580, p<0.05) association in T1. SOD (r=0.445) and CAT (r=0.949) had a highly significant (p<0.05) correlation with sperm motility in T1. No internalization of ZnO and Se NPs were observed under TEM. HSP70 and HSP90 mRNA levels were significantly (p<0.001) higher in T1 in comparison to other groups. HSP70 and HSP90 expression levels had a significant (p<0.05) positive correlation with motility in group T1. No significant (p>0.05) differences in pregnancy rates following AI were recorded among the different treatment groups in comparison to control. In conclusion, extender supplemented with 0.1 mg/mL ZnO NPs improved post-thaw semen quality of cryopreserved Assam Hill goat spermatozoa consequently by lowering lipid peroxidation and increasing expression of cryostress associated heat shock genes.
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    ThesisItemOpen Access
    ULTRASONOGRAPHIC MONITORING OF OVARIAN FOLLICULAR AND LUTEAL DYNAMICS IN COW
    (College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2022-03) RALTE, VANLALNGILNEII; Dutta, Devojyoti
    The ovarian follicular and luteal dynamics, blood flow pattern in the corpus luteum (CL) including the pattern of follicular wave and luteal development were studied using portable ultrasound device with 5-10 MHz linear array transrectal probe in female Crossbred and Lakhimi, indigenous cattle breed of Assam. In the experiment, twelve each cyclic Crossbred (Jersey X L) and Lakhimi (L) cows were subdivided into two groups viz. Group-I/Natural or Spontaneous oestrus and Group-II/Induced or Synchronized oestrus for each breed comprising of six cows in each group. They were evaluated in the subsequent cycle of natural luteolysis (Group-I) and induced luteolysis by prostaglandin F2α (Group-II) and circulating steroids were estimated on alternate days of the cycle. From scanning the ovaries for two inter-ovulatory cycle the experimental cows exhibited two and three follicular waves per cycle. Most of the cows 30(62.50 %) exhibited 3-wave cycle and 18(37.50 %) had 2-wave cycle. The diameter of 12.4 mm and 10.5 mm were the threshold diameter for ovulation according to follicle diameter in crossbred and Lakhimi cows, respectively. The inter-ovulatory interval (IOI) was found to be significantly longer (P<0.05) in Crossbred cows experiencing 3-wave (21-22 day) than 2-wave (20-21 day) per cycle. The first wave emerged on day 0 (day of ovulation) to day 1 of cycle with mean day of 0.13 ± 0.12 to 0.83 ± 0.38 in both the breed and group. The second wave onset was significantly (P < 0.05) earlier (9.00 ± 0.19, 8.50 ± 0.22) in 3-wave cycle than the 2-wave cycle (10.50 ± 0.29, 10.67 ± 0.21) in both Crossbred and Lakhimi cows. The average number of follicles (≥ 2 mm) observed on wave onset was 10-13 in Crossbred and 9-13 in Lakhimi cows. The first wave dominant follicle (DF) became deviated in between day 3 to 4 in both the breeds, while the second wave DF deviated on day 11-12 in Crossbred and day 10-11 in Lakhimi cows exhibiting 2-wave cycle. Deviation of the third DF occurred on day 17-18 in both the breeds. Maximum mean diameter of DF in the second waves recorded as 8.74 ± 0.52 and 7.70 ± 0.27 mm in Crossbred and Lakhimi cows respectively in 3-wave cycles was significantly smaller (P < 0.05) than the second wave DF diameter (12.83 ± 0.65 and 10.53 ± 0.76 mm) in 2-wave cycles in both crossbred and Lakhimi cows. The DF maximum diameter was attained significantly (P < 0.05) earlier in 3-wave cycle in both the first and second wave than in 2-wave cycle. The average growth rate of ovulatory DF in 3-wave cycle was 1.19 and 1.37 mm/day in Group-I and II respectively in Crossbred while, 0.88 and 1.18 mm/day in Group-I and II respectively in Lakhimi. The dominant follicles of the first wave (non-ovulatory) began atresia on day 8.0 ± 0.41 at the rate of 1.36 to 1.53 mm/day in Crossbred cow and on day 7.75 ± 0.25 at the rate of 0.97 mm/day in Lakhimi cows. The developing CL was detected on day 0 to day 1 in both Crossbred and Lakhimi cows. Maximum diameter (mm) of the CL for the two and three follicular wave cycle in Lakhimi (15.29 ± 0.64 mm and 15.08 ± 0.45 mm) were significantly smaller (P <0.05) than in Crossbred cows (22.98 ± 0.87 mm and 21.94 ± 0.66 mm). Onset of luteal regression of the 2-wave cycles (day 12 to 13 ) was significantly earlier (P < 0.05) than in the 3-wave cycle (day 15 to 16 ). In early luteal phase (day 0 to 5) the Doppler signal increased to 65.73 ± 5.02 mm2 in Crossbred and 57.30 ± 9.83 mm2 in Lakhimi. During the mid-luteal phase (day 7 to 13) the area gradually increased to 119.82 ± 5.54 mm2 in Crossbred and 93.82 ± 4.12 mm2 in Lakhimi cows then rapidly declined in the late luteal phase (day 15 to 18) to 10.66 ± 2.02 mm2 in Crossbred and 11.99 ± 5.38 mm2 in Lakhimi. Similar pattern was observed in 2-wave cycle. On the day of oestrus, the mean serum Oestradiol-17β level ranged from 35.75 ± 0.64 to 51.01 ± 2.13 pg/ml in Crossbred and 32.43 ± 0.74 to 35.74 ± 0.97 pg/ml in Lakhimi while the serum progesterone level ranged from 0.58 ± 0.71 to 1.27 ± 0.33 ng/ml in crossbred and 0.43 ± 0.77 to 0.47 ± 0.76 ng/ml in Lakhimi cows. ii There was a positive correlation between LBF area and progesterone level in 2-wave crossbred (r = 0.92), 2-wave Lakhimi (r= 0.82), 3-wave crossbred (r = 0.81) and 3-wave Lakhimi (r = 0.90) during the cycle. There was positive correlation between DF size and oestradiol level in 2-wave (r = 0.40) also in 3-wave crossbred and Lakhimi cows (r = 0.41). Negative correlation was observed between DF size and progesterone level in 2-wave (r = - 0.47) and in 3-wave (r = - 0.40) in crossbred and Lakhimi cows.