Thesis Abstract and Dissertation Abstracts
Cryopreservation Of Bovine Semen With Some Additives For Augmenting Fertility
Abstract Category: Science
Course / Degree: MVSc
Institution / University: Orissa University of Agriculture and Technology, Bhubaneswar, India
Published in: 2008
The present experiment was under taken in the Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Bhubaneswar in collaboration with Frozen Semen Bank, Cuttack. The objective of the study was to determine the efficacy of Cysteine Hydrochloride and Glutathione as additives in routine frozen semen extender (EYTG) at the rate of 5 mM.
Various seminal traits i.e. volume, mass activity, individual motility, sperm concentration, total sperm abnormality, loss of acrosomal integrity and HOST were analyzed in extended semen containing cysteine (group II), or glutathione (group III) with routine control (group I). Post thaw seminal attributes relating to motility, livability, total sperm abnormality, loss of acrosomal integrity, cervical mucus penetration test, estimation of MDA concentration, hypo-osmotic swelling positive sperm per cent and mitochondrial membrane potential DNA fragmentation test were assessed in various experimental groups. Conception rate of various experimental groups were evaluated on the basis of field fertility trial.
Six ejaculates of each good and poor freezable crossbred bull were collected. The average volume was 6.40 ± 0.41 ml and 5.20 ± 0.36 ml respectively in good and poor freezable crossbred bull. The mass activity of good and poor freezable cross bred bulls was more or less similar with figures of 2.50 ± 0.13 and 2.40 ± 0.13, respectively. The average sperm concentration (X 106) per ml was 1240.26 ±82.77 and 1007.53 ±50.28 in good and poor freezable crossbred bull semen respectively. The corresponding values for motility percent were 77.13± 0.60 and 75.33± 0.77 respectively. On an average, the live sperm per cent was more than 85% for both good and poor freezable semen. The per cent of total abnormal sperm was 6.33 ±0.19 and 9.39 ± 0.27 respectively in good and poor freezable bulls. The loss of acrosomal integrity per cent was higher (12.73 ±1.12) in poor and lower (6.46 ± 0.90) in good freezable bulls. The hypo-osmotic swelling positive spermatozoa per cent was higher (85.78 ±0.42) in good and lower (80.38 ±0.32) in poor freezable bulls. The seminal parameters like individual motility, live sperm percent, total sperm abnormality, loss of acrosomal integrity and HOST showed highly significant difference (p < 0.01) between the groups. The other parameters like volume and sperm concentration also differed significantly (p < 0.05).
The individual motility of post freeze semen showed more than 50% overall value irrespective of extender used, for all the experimental bulls. The good freezable crossbred bull had showed higher motility percent than poor freezable crossbred bulls, irrespective of additives used. But the glutathione added group showed highest motility of 74.12 ± 0.39 and it was lowest (61.83 ± 0.48) in cysteine hydrochloride added semen in good freezable bulls. Similarly the glutathione extended semen also showed same trend in poor freezable bull (66.83 ±0.42). The overall average motility per cent of group I, II and III were also more than 50%. Comparison of individual motility value in experimental groups recorded highly significant difference (p < 0.01) among three groups for both the good and poor freezable crossbred bulls.
The post freeze live sperm percent was observed in glutathione (group III) and cysteine (group II) treated group of good freezer with the value of 76.28 ± 0.37 and 70.85 ± 0.32 while in control group (group I), it was 63.09 ± 0.37. In the poor freezer, it was 69.32 ± 0.35, 60.92 ± 0.89 and 54.32 ± 0.24, respectively. Analysis of variance revealed a high significant difference (p < 0.01) in treatment and control groups in both good and poor freezable semen.
The total sperm abnormality percent in good freezer revealed almost similar value for group III (9.45 ± 0.94) followed by group II (11.42 ± 0.79) and group I (13.18 ± 1.41). The similar trend was found in poor freezer with a value of 14.26 ± 0.18, 15.87 ± 0.26 and 18.06 ± 0.18 respectively while both the classes showed highly significant difference (p < 0.01) among groups.
The loss of acrosomal integrity for group III was 8.44 ± 0.63 and 13.01 ± 0.60 for good and poor freezer respectively, which was significantly different (p < 0.01) from rest of the groups in respective category. On the contrary, group II and group I value did not differ significantly.
The post thaw value of HOST per cent in good freezer bulls was 65.74 ± 0.77, 70.47 ± 0.35 and 81.19 ± 0.30 in group I, II and III respectively. The corresponding values for poor freezable bulls were 57.03 ± 0.32, 57.65 ±0.47 and 71.29 ± 0.26, respectively. Exhibition of highly significant difference ( p < 0.01) was observed among groups in both good and poor freezers. The glutathione added group showed significantly higher rating for HOST with respect to freezing type.
The vanguard distance (mm) travelled by the sperm in bovine cervical mucus after thawing was higher in glutathione treated group than the other group in both good (31.16 ± 1.09) and poor (25.44 ± 1.35) freezable crossbred bulls. The cysteine hydrochloride treated group revealed second highest value next to the former in both good (27.76 ± 0.98) and poor (23.35 ± 1.18) freezable bulls. The control untreated group recorded lowest value (25.84 ± 1.35 and 20.24 ± 1.09) as compared to former groups.
DNA fragmentation of post thawed semen revealed that untreated control (10.77 ± 0.67 and 24.70 ± 1.16) group had higher percent of fragmentation in both good and poor freezer as compared to cysteine ( 8.83 ± 1.13 and 16.42 ± 2.19) and glutathione (6.94 ± 1.28 and 9.98 ± 1.17) treated groups.
The MDA production (nmol/108 sperms) of post thaw semen in group III (2.17 ± 0.82 and 3.17 ± 1.34) had lower value in both good and poor freezable crossbred bull as compared to group II (2.60 ± 0.76 and 3.50 ± 1.42) and control (2.73 ± 0.85 and 3.68 ± 1.65).
The percent of mitochondrial membrane potential positive sperm of post thaw semen revealed that glutathione (29.78 ± 0.98 and 27.55 ± 1.64) treated group had higher value in both good and poor freezable crossbred bull as compared to cysteine (28.63 ± 3.08 and 26.11 ± 2.96) treated and control (26.11 ± 3.27 and 25.93 ± 1.60 ).
Assessment of motility at different hours of incubation in good and poor freezable crossbred bull semen by CASA revealed that the forward progressive motility was higher in good freezer from zero to four hours of incubation than poor freezer. Analysis of variance has also revealed that highly significant difference (p < 0.01) existing at 1st hour and significance difference at 0, 2 and 4 hours of incubation. With regards to non- progressive motility the poor freezer found showing value similar to good freezer at all hours of incubation. Analysis of variance had revealed that highly significant (p < 0.01) difference is existing at 1st hour and no significance difference at 0, 2 and 4 hours of incubation. Assessment of total motility have also behaved like forward progressive motility. The good freezer have higher value as compared to poor freezer at all hours of incubation period. Analysis of variance had revealed significance difference (p < 0.05) at 4th hour and no significance difference at 0, 1 and 2 hours of incubation. As compared to different hours of incubation, the forward progressive motility and total motility have been lowered from 0 hours to 4 hours, but the non- progressive motility and static sperm percent were found increasing gradually from 0 hours to 4 hours of incubation.
Comparison of motility between control and treatment groups at different hours of incubation has revealed that the forward progressive motility, non – progressive motility and total motility was higher in glutathione treated group at different hours of incubation period than cysteine treated and untreated control group. Contrastingly the total static sperm percent was higher in control group than either of glutathione or cysteine treated group. Analysis of variance revealed that the forward progressive motility, non – progressive motility and total motility were showing highly significant difference (p < 0.01) at 1 hour incubation period and there was no significant difference at 0, 2 and 4 hours incubation periods. Total static sperms was not showing any significant difference at different hours of incubation.
Computer assisted semen analysis (CASA) of some post-thaw seminal characteristics in control and treatmentgroups at different hours of incubation has revealed that the CASA parameters like curvilinear velocity, straight line velocity, average path velocity, linearity, straightness, wobble, amplitude of lateral head displacement and beat/cross frequency were higher in good freezer as compared to poor freezer at different hours of incubation and glutathione have higher value than other treatment and control groups in both the good and poor freezable crossbred bulls at different hours of incubation.
Analysis of variance of the different CASA parameters revealed that there was no significant difference between the experimental groups at different hours of incubation except curvilinear velocity and amplitude of lateral head displacement at 0 hours, which was showing highly significant difference (p < 0.01).
The post insemination conception rate in various semen additives showed that the glutathione treated extender has registered highest no. of pregnancies to the tune of 72.00% and 59.00% respectively in good and poor freezable crossbred bulls out of 51 and 47 artificial inseminations. Similarly the overall pregnancy rate was 60.00% and 56.00 % in cysteine added frozen semen, out of 41 and 44 artificial breeding. The lowest value of 51.00% and 48.00% was observed in control group where no additives were used in a routine frozen semen extender (EYTG). The glutathione treated group recorded a highest conception rate of 66.00% and it was lowest (49.00%) for control untreated group. Comparison of conception rate between experimental groups revealed no significant difference by chi-square analysis. Similarly the pregnancy percentages registered by each bull did not differ significantly and the overall value between applications also was not significant, but glutathione had higher percentage of pregnancy.
Thesis Keywords/Search Tags:
Reduced Glutathione, Cysteine Hydrochloride, Computer assisted sperm analysis, Mitochondrial membrane potential, FITC-PSA
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Submission Details: Thesis Abstract submitted by perumal ponraj from India on 25-May-2013 09:01.
Abstract has been viewed 2623 times (since 7 Mar 2010).
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