Please use this identifier to cite or link to this item: http://krishikosh.egranth.ac.in/handle/1/5810139728
Authors: M. RAJASHRI
Advisor: Dr. K. CHANDRASHEKAR REDDY
Title: STUDIES ON ASSESSMENT OF POST THAW SEMEN QUALITY AND FERTILITY OF MURRAH BUFFALO BULLS
Publisher: PVNR TVU
Language: en
Type: Thesis
Pages: 179
Agrotags: null
Abstract: The present investigation was undertaken to study the seminal characteristics of frozen thawed semen of Murrah buffalo bulls (aged 3-6 years) maintained at the Frozen semen bull station (FSBS), Karimnagar (18.4386° N, 79.1288° E) and to evaluate the relationship between the evaluated seminal attributes and the in vivo fertility. Experiment was designed to study the semen characteristics immediately after semen collection (Semen volume, sperm concentration and sperm individual motility) and comparison of post-thaw semen quality was done by assessment of Sperm individual motility, Live spermatozoa percentage, Plasma membrane integrity by HOST, Sperm abnormalities, Acrosomal integrity (Giemsa, FITC – PSA and Coomassie brilliant blue Staining), DNA integrity by Toluidine Blue Staining, Immature sperm percentage by Aniline Blue Staining, Mitochondrial activity (Diaminobenzidine and Rhodamine 123 staining). The average semen ejaculate volume, sperm concentration and sperm individual motility percentage of Murrah Buffalo bull semen were 2.52 ± 0.09 ml, 880.10 ± 22.71 and 78.02 ± 0.84. Semen ejaculate volume showed low positive correlation with semen concentration (r = 0.14) and no correlation with sperm individual motility (r = 0.09) while semen concentration showed significant positive correlation with sperm individual motility (r = 0.18*). A total of number of 588 semen straws (42 straws/each bull) were utilized for semen evaluation in the present study. Selected Murrah buffalo bulls (14) were categorized into high (n=7) and low (n=7) fertility bulls as per the records of FSBS. The overall post thaw motility percentage of Murrah Buffalo bull semen recorded was 50.59 ± 0.45 while high fertility and low fertility bull groups recorded were 50.74 ± 0.31 and 50.40 ± 1.35, respectively. The overall live spermatozoa xxiii percentage of Murrah Buffalo bull semen recorded was 62.42 ± 0.85 while high fertility and low fertility bull groups recorded were 65.28 ± 1.03 and 59.63 ± 1.31, respectively. Higher sperm livability percentage was recorded in high fertility buffalo bull group (65.28%) compared to that of low fertility buffalo bull group (59.63%). The overall mean percentage of Murrah buffalo bull sperm positive to HOS test (150 and 100 mosm/L) was 41.60 ± 0.99 and 47.98 ± 1.11. High fertility bull group recorded 44.07 ± 1.30 and 50.83 ± 1.38 percentage while low fertility bull group recorded 39.10 ± 1.45 and 45.44 ± 1.66 percentage of sperm positive to HOS test (at 150 and 100 mosm/L), respectively. The overall mean sperm abnormalities percentage of Murrah Buffalo bull semen recorded was 8.44 ± 0.34 while high fertility and low fertility bull groups recorded were 8.29 ± 0.46 and 8.57 ± 0.50, respectively. The mean sperm acrosomal integrity percentage of Murrah Buffalo bull semen recorded by giemsa staining, FITC-PSA staining and Coomassie brilliant blue staining were 63.82 ± 0.98, 65.58 ± 0.71 and 69.36 ± 0.85, respectively. The mean acrosomal integrity percentage recorded for high fertility and low fertility bull groups by giemsa staining were 65.08 ± 1.06 and 62.64 ± 1.61, FITC-PSA staining were 66.00 ± 0.87 and 65.20 ± 1.11 and Coomassie brilliant blue staining were 69.73 ± 1.18 and 69.02 ± 1.22, respectively. The overall mean sperm DNA integrity percentage of Murrah Buffalo bull semen assessed by Toluidine blue staining was 98.54 ± 0.09 while high fertility and low fertility bull groups recorded were 98.58 ± 0.13 and 98.50 ± 0.14, respectively. The overall mean immature spermatozoa percentage of Murrah Buffalo bull semen assessed by Aniline blue staining was 1.27 ± 0.02 while high fertility and low fertility bull groups recorded were 1.23 ± 0.02 and 1.32 ± 0.02, respectively. The overall mean mitochondrial activity percentage obtained in Class I, II, III and IV of Diaminobenzidine (DAB) staining in Murrah Buffalo bull semen were 18.95 ± 1.01, 37.80 ± 1.12, 29.32 ± 0.96 and 13.92 ± 0.90, respectively while high fertility bull group recorded 22.79 ± 1.56, 34.71 ± 1.50, 29.38 ± 1.38 and 13.10 ± 1.16, and low fertility bull group recorded 15.38 ± 1.06, 40.66 ± 1.55, 29.26 ± 1.35 and 14.69 ± 1.37 percentage, respectively. The overall mean percentage of sperm positive for mitochondrial activity assessed by Rhodamine 123 staining in Murrah Buffalo bull semen was 62.33 ± 0.75 while high fertility and low fertility bull groups recorded were 66.30 ± 0.87 and 58.62 ± 0.93, respectively. A total of 490 buffaloes (Number of inseminations/bull=35) in proper stage of estrus and free from uterine infections were inseminated by using the same batch of frozen semen used for post thaw semen evaluation. The insemination was carried out in five regions in and around regions of Karimnagar district (Karimnagar, Eligaidu, Odela & Ramagundam) and Warangal district (Gavicherla village). Pregnancy was confirmed by per-rectal palpation after 60 days of AI in non returned buffaloes and the conception rates were estimated. The relationship between the evaluated seminal attributes with the in vivo fertility was assessed. The overall mean conception rate percentage recorded for Murrah Buffalo bull semen was 53.04 ± 2.60 while high fertility and low fertility bull groups recorded were xxiv 60.20 ± 1.90 and 45.88 ± 2.96, respectively. Higher percentage of sperm livability, HOS test (at 150 and 100 mosm/L), mitochondrial activity (Diaminobenzidine and Rhodamine 123 staining) and conception rates were recorded in high fertility buffalo bull group compared to that of low fertility buffalo bull group. There was no significant difference (P≥0.05) observed between the groups pertaining to post thaw motility, sperm abnormalities, acrosomal integrity (Giemsa, FITC – PSA and Coomassie brilliant blue Staining), DNA integrity and Immature spermatozoa obtained per ejaculation while significant difference (P≤0.05) was observed between high fertility group and low fertility group buffalo bulls pertaining to sperm livability and HOS test (at 150 and 100 mosm/L), Mitochondrial activity (Diaminobenzidine and Rhodamine 123 staining) and conception rates. Significant difference (P≤0.05) was observed between individual bulls within the groups pertaining to post thaw motility, sperm livability, HOS test (100 mosm/L), sperm abnormalities, acrosomal integrity (Giemsa, FITC – PSA and Coomassie brilliant blue Staining) and Mitochondrial activity (Diaminobenzidine and Rhodamine 123 staining) while there was no significant difference between individual bulls within the groups pertaining to HOS test (at 150 mosm/L), DNA integrity and Immature spermatozoa percentage. Conception rate showed low correlation with post thaw motility (PTM) (r = 0.19), Hypo-osmotic Swelling Test (HOST-100mOsml/) (r = 0.41), DNA integrity (r = 0.19) and FITC-PSA Staining (r = 0.39) while significant positive correlation was observed with Live Spermatozoa Percentage (r = 0.58*), Hypo-osmotic Swelling Test (HOST-150mOsml/) (r = 0.54*), Giemsa staining (r = 0.55*), DAB I staining (r = 0.27*) and Rhodamine staining (r = 382**). Conception rate did not show correlation with sperm individual motility (r = 0.01), sperm abnormalities (r = 0.00), Coomassie brilliant blue staining (r = 0.09), DAB III staining (r = -0.08) and DAB IV staining (r = 0.07) while low negative correlation was observed with semen ejaculate volume (r = -0.11), semen concentration (r = -0.12), Immature sperm percentage (r = -0.28) and DAB II staining (r = -0.24*). Post thaw motility showed no correlation with livability (r = 0.00) and Hypo-osmotic Swelling Test (HOST-100 & 150mOsml/) (r = 0.02; r = 0.02) and immature sperm percentage (r = 0.04), respectively. Low negative correlation was observed between PTM and sperm abnormalities (r = -0.17) while low positive correlation was observed with DNA integrity percentage (r = 0.20). Livability showed significant positive correlation with Hypo-osmotic Swelling Test (HOST-100 & 150 mOsml/) (r = 0.65* & 0.52*) while no correlation was observed with sperm abnormalities (r = -0.057), DNA integrity (r = 0.00) and Immature sperm percentage (r = 0.00) respectively. HOST (150 mOsm/L) showed significant high positive correlation with HOST (100 mOsm/L) (r = 94**) and no correlation was observed with sperm abnormalities (r = 0.08) while low positive correlation was observed with DNA integrity (r = 0.41). Low negative correlation was observed with HOST (150 mOsm/L) and Immature sperm percentage (r = -0.31). Sperm abnormalities percentage did not show any correlation with DNA integrity percentage (r = -0.02) and immature sperm percentage (r = 0.09) while DNA integrity percentage showed negative correlation with that of immature sperm percentage (r = -0.51). Giemsa staining showed significant high xxv positive correlation with FITC-PSA staining (r = 0.85**) and moderate positive correlation with Coomassie brilliant blue staining (r = 0.50) while FITC-PSA staining showed low positive correlation with Coomassie brilliant blue staining (r = 0.42). The present investigation revealed that the seminal characteristics of fresh and frozen thawed semen obtained from the Murrah buffalo bulls of FSBS were within the normal acceptable limits. Among two experimental groups, high fertility bull group showed significantly (p<0.05) higher sperm livability, HOS test (at 150 and 100 mOsm/L), mitochondrial activity (Diaminobenzidine and Rhodamine 123 staining) and conception rates compared to low fertility bull group. From above observation, it was observed that the tests that could predict buffalo bull fertility with high accuracy included liveability, Hypo-osmotic swelling test, Mitochondrial activity assay by Rhodamine staining and DAB staining which also showed correlation with the conception rate. Thus, it was concluded that these tests may be recommended for analysis of post thaw semen quality at field level for evaluating the fertilizing ability of Murrah Buffalo bulls.
Description: D-683
Subject: Veterinary Gynaecology and Obstetrics
Theme: VGO
These Type: Ph.D
Issue Date: 2019-12-27
Appears in Collections:Thesis

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