1.
Effect Of Two Synchronization Protocols (Ovsynch Va 2Pg)And Gnrh Pn Day Of Breeding On Reproductive Performancein
by Hasan Riaz | Prof. Dr. Nasim Ahmad | Dr. Mian Abdul Sattar | Prof. Dr.
Material type: ; Format:
print
Publisher: 2011Dissertation note: The main objective of the present study is to compare the two synchronization protocols (Ovsynch vs 2PG) on estrus response, follicular development, ovulation, pregnancy rate and fecundity in Beetal and Teddy breeds of Pakistan. The secondary objective is to determine the efficacy of GnRH at the time of breeding on pregnancy rate in Beetal goats. Experiment 1 was conducted with a total of 28 goats during breeding season (September-October) at Livestock Production Research Institute, Bahadurnagar, Okara. The goats were randomly allocated in one of two treatment groups, Ovsynch and two PGF treatments. The Ovsynch treated does received an intramuscular injection of GnRH analogue 12.5µg lecirelin (Dalmarelin®, Fatro, Italy), on "Day 0". On "Day 7" does were treated with injection of PGF2? analogue 37.5µg d-cloprostenol (Dalmazin®, Fatro, Italy,) followed by a second injection of 12.5µg lecirelin (Dalmarelin®, Fatro, Italy), after a 48h period. Animals in the double PGF group received two injections of 37.5µg d-cloprostenol (Dalmazin®, Fatro, Italy), 10 days apart. In order to determine the estrous response and its duration, two aproned bucks were used twice daily for 30 minutes after the last PGF2? injection in both groups until the termination of estrous signs. All goats were naturally bred 12h after the onset of estrus. Follicular and luteal activity was monitored by transrectal Ultrasonography (ALOKA, SSD 900 Japan) with the aid of a 7.5-MHz linear array transducer daily- from the start of treatment, until estrus and then twice a day until the cessation of large follicles (apparent ovulation). On day 7 after breeding, all goats were re-examined by transrectal ultrasonography to determine the ovulation rate, based upon the number of CL's present. Pregnancy rate and fecundity (no of embryos per doe) was again determined at Day 30 after breeding by transrectal ultrasonography. Experiment 2 was performed on 25 goats. These does were randomly allocated to two groups (GnRH, n=11 and Control, n=14). GnRH does received 12.5µg lecirelin on the day of natural breeding (12h after standing estrus), while the control does did not receive any treatment. Timing of ovulation, pregnancy rate and fecundity were assessed by transrectal ultrasonography as in Experiment 1. Estrus (Experiment 1) was detected in 10 out of 14 does (71%) in Ovsynch group, while all 14 goats (100%) of double PGF responded group. The interval from the last PGF injection to estrus was 48.0 ± 2.6h in the Ovsynch group, while 36.0 ± 1.2h in the double PGF group (P<0.05). The interval from the last PGF injection to ovulation did not differ significantly between the Ovsynch (73.0 ± 4.8h) and double PGF (66.0 ± 2.7h) groups respectively. Similarly the interval from standing estrus to ovulation was not significant between the Ovsynch (24.0 ± 3.7h) and double PGF (30.0 ± 2.7h) treatments respectively. Estrous duration was similar in both treatment groups (44.7 ± 4.9h for Ovsynch and 47.1 ± 2.9h for the 2PGF). The size of ovulatory follicle was similar in both treatment groups (Ovsynch, 7.1 ± 0.2mm; 2PGF, 7.1 ± 0.1mm). On day 7 after breeding, the ovulation rate (no. of CL's on the ovary) were similar in both the Ovsynch (1.6 ± 0.2) and double PGF (2.0 ± 0.2) groups. The pregnancy rate as determined by transrectal ultrasonography on day 30 (Fig. 1) revealed 6/10 (60%) in the Ovsynch and 11/14 (78%) in the double PGF groups respectively (not significant). Fecundity was similar for both treatment groups (1.6 ± 0.2). In Experiment 2 the interval from standing estrus to ovulation recorded was not significant (31.2 ± 2.9h and 36.0 ± 5.3h), the ovulatory diameter (6.8 ± 0.1mm, 7.2 ± 0.5 mm (P<0.05), the pregnancy rate (54% (6/11) and 64% (9/14), and fecundity (1.5 ± 0.5, 1.7 ± 0.5) were not significant between GnRH treated and control does respectively (Table 2). From these experiments it can be concluded that the Ovsynch protocol, which is a non-steroidal and safe approach for estrous synchronization appears to be similar to the double PGF injection protocol, in terms of the reproductive performance in goats. However, this needs to be tested on larger number of animals. Furthermore, the use of GnRH at the time of breeding does not improve the reproductive performance in goats. The data generated has important implications for commercial goat farming in the developing countries.
Availability: Items available for loan: UVAS Library [Call number: 1295,T] (1).
2.
Effect Of Cholesterol Addition To Buffalo Bull Semen During Cryopreservation Procedures On Post-Thaw Survival Of Spermatozoa
by Ahmad Yar Qamar | Dr. Mian Abdul Sattar | Prof. Dr | Prof. Dr. Nasim Ahmad.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Cholesterol is an important constituent in the membrane fluidity regulation. This role becomes
more important in the cryopreservation of sperm cells where destabilization of plasma membrane
leads to intracellular ice formation causing death of the cell. The objective of the present study
was to verify if addition of cholesterol in semen extender has a beneficial effect on post-thaw
semen quality in buffalo bulls. This study was carried out at Al-Haiwan Sires, Sahiwal, Pakistan.
Cholesterol was added to Tris-citric acid semen extender in the form of cholesterol-loaded
cyclodextrin (CLC). Semen was collected by using artificial vagina (420 C) from four buffalo
bulls. Split pooled ejaculate (n = 7), having more than 60 % visual sperm motility, were diluted
at 370 C in extender containing CLC either 3mg Iml (LOW), 4 mgl ml (MED), 5 mg Iml (HIGH)
or without (CON). Motility of semen samples before freezing was checked and cryopreservation
was done through routine procedures. Beneficial effect of CLC in cryopreserved semen was .
assessed by post-thaw motility (%), live spermatozoa (%), morphological abnormalities (%),
normal acrosornes (%) and plasma membrane integrity (%) in four groups. Analysis of variance
revealed that motility before freezing was significantly (P<0.05) more in MED (68.93 ± 0.51 %)
as compared to CON and HIGH but non-significantly higher than LOW. Percent post thaw
motility (PTM) from semen samples containing MED concentration of CLC was the highest
(61.43 ± 0.74 %; P < 0.05) followed by LOW (50.36 ± l.15 %; P < 0.05) or HIGH (38.57 ± 2.66
%; P < 0.05) (Table 1). Percent PTM was the lowest in CON (32.86 ± 2.07 %; P < 0.05) semen
samples. Mean plasma membrane integrity (PM I) from semen samples containing MED
concentration of CLC was the highest (59.36 ± 1.32 %; P < 0.05) followed by LOW (51.93 ±
1.32 %; P < 0.05) or HIGH (44.00 ± 1.65 %; P < 0.05) and was least in CON (41.14 ± 1.71 %; P
< 0.05) semen samples. Mean normal acrosomes from semen samples containing MED
concentration of CLC was the highest (52.93 ± 1.78 %; P < 0.05) followed by HIGH (40.57 ±
2.01 %; P < 0.05) and CON (34.93 ± 2.89 %; P < 0.05) semen samples. Buffalo bull semen in
MED & LOW had significantly (P < 0.05) higher live sperm percentage (69.64 ± 1.84 & 61.43 ±
1.62, respectively) as compared to HIGH and CON samples. CON samples had significantly
(P<0.05) higher morphological abnormalities (9.86 ± 0.14 %) as compared to LOW & MED but
non-significantly higher as compared to HIGH samples. All the variables under study had strong
positive correlation (P < 0.01) with each other except morphological abnormalities having strong
negative correlation (P < 0.01) with all others. It is concluded that addition of cholesterol in
MED concentration (4 mg CLCI ml of semen extender) to buffalo bull semen can improve post-
thaw semen quality.
Conception rate of AI in buffaloes is generally lowered than in cows. Cholesterol is an
important constituent in the membrane fluidity regulation. This role becomes more important in
the cryopreservation of sperm cells where destabilization of plasma membrane leads to
intracellular ice formation causing death of the cell. The objective of the present study was to
verify if addition of cholesterol in semen extender has a beneficial effect on semen post-thaw
quality in buffalo bulls. This study was carried out at Al-Haiwan Sires, Sahiwal, Pakistan.
Cholesterol was added to Tris-citric acid semen extender in the form of cholesterol-loaded
cyclodextrin (CLC). Semen was collected by using artificial vagina (42° C) from four buffalo
bulls. Split pooled ejaculate (n = 7), having more than 60 % visual sperm motility, were diluted
at 37° C in extender containing CLC either 3mg Iml (LOW), 4 mgl ml (MED), 5 mg Iml (HIGH)
or without (CON). Motility of semen samples before freezing was checked and cryopreservation
was done through routine procedures. Further examination was done to check post-thaw motility
(%), live spermatozoa (%), morphological abnormalities (%), normal acrosomes (%) and plasma
membrane integrity (%) using phase contrast microscope and hypo osmotic swelling assays.
Analysis of variance revealed that motility before freezing was significantly (P<0.05) more in
MED (68.93 ± 0.5 I %) as compared to CON and HIGH but non-significantly higher than LOW.
Percent post thaw motility (PTM) from semen samples containing MED concentration of CLC
was the highest (61.43 ± 0.74 %; P < 0.05) followed by LOW (50.36 ± 1.15 %; P < 0.05) or
HIGH (38.57 ± 2.66 %; P < 0.05) (Table 1). Percent PTM was the lowest in CON (32.86 ± 2.07
%; P < 0.05) semen samples. Mean plasma membrane integrity (PMI) from semen samples
containing MED concentration of CLC was the highest (59.36 ± 1.32 %; P < 0.05) followed by
LOW (51.93 ± 1.32 %; P < 0.05) or HIGH (44.00 ± 1.65 %; P < 0.05) and was least in CON
(41. I 4 ± 1.71 %; P < 0.05) semen samples. Mean normal acrosomes from semen samples
containing MED concentration of CLC was the highest (52.93 ± 1.78 %; P < 0.05) followed by
HIGH (40.57 ± 2.01 %; P < 0.05) and CON (34.93 ± 2.89 %; P < 0.05) semen samples. Buffalo
bull semen in MED & LOW had significantly (P < 0.05) higher live sperm percentage (69.64 ±
1.84 & 6 1.43 ± 1.62, respectively) as compared to HIGH and CON samples. CON samples had
significantly (P<0.05) higher morphological abnormalities (9.86 ± 0.14 %) as compared to LOW
& MED but non-significantly higher as compared to HIGH samples. All the variables under
study had strong positive correlation (P < 0.01) with each other except morphological
abnormalities having strong negative correlation (P < 0.01) with all others. It is concluded that
addition of cholesterol in MED concentration (4 mg CLCf ml of semen extender) to buffalo bull
semen can improve post-thaw semen quality.
Availability: Items available for loan: UVAS Library [Call number: 1398,T] (1).
3.
Effect Of (Estradiol Benzoate) On Estrus Response And Fertility In Cidr Treated Crossbred Heifers.
by Muhammad Usman Mehmood | Dr. Mian Abdul Sattar | Dr. Muhammad | Prof. Dr. Nasim Ahmad.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Commercial dairy farming in Pakistan is rapidly growing. Crossbred dairy animals hold promise because of two reasons, first that they are significantly high producer than buffalo, Sahiwal, Cholistani and Desi cows and secondly that they are much more adaptable as compared to exotic (Friesian or Jerseys) cows. Generally, replacement heifers remain ignored by the owners in term of feeding and early achievement of puberty as they are not producing. Estradiol promotes tighter synchrony and ovulation and provides opportunity for fixed time A.I as reported earlier in Holstein dairy cows. Use of CIDR has been reported to be effective for synchronization in variety of dairy animals including buffaloes, dairy and beef cows. However no such report has been published on crossbred heifers. The objectives of the present study were to investigate the effect of estradiol benzoate (EB) on estrus intensity, response and fertility in CIDR treated cross bred heifers.Crossbred heifers with average body condition of 3 ± 0.5, located at four farms were assigned throughout one year to receive one of the two treatments. Crossbred heifers with EB (n = 50) or without EB (n = 50). All the heifers were treated with Controlled Internal Drug Releasing Device (CIDR; 1.38 mg Progesterone; Pfizer, USA) on day 0 and were administered injection of PGF2?, (0.150 mg; Dalmazine®, Fatro, Italy; 2 ml; i.m.) on day 6 followed by removal of CIDR on day 7. Estradiol benzoate (400µg/4ml, i.m. Sigma Aldrich Germany) was administered 24 h after the CIDR removal. Estrus detection was carried out by visual observation; twice daily for at least 30 min, beginning after CIDR removal till insemination. All heifers were fixed time inseminated with frozen-thawed semen after 48 and 60 h after CIDR removal, respectively. Pregnancy status was determined by ultrasonography (HS 1500V; Honda; Japan, with 7.5 MHz probe) between days 30 and 40 post AI. Estrus response and pregnancy rate were analyzed using Chi-square test. Pregnancy rate was determined by number pregnant/number treated x 100. The estrus response was 100% in both the groups. The estrus intensity was higher 2.9 ± 0.1in the EB heifers compared to the controls 2.0 ± 0.7. The results showed that pregnancy rate was 54% (27/50) in EB and 36% (18/50) in without EB heifers. The overall pregnancy rate was 45% (45/100). It is concluded that estradiol benzoate in CIDR protocol intensified the signs of estrus without affecting the response, with higher fertility in crossbred heifers.
Availability: Items available for loan: UVAS Library [Call number: 1502,T] (1).
4.
Reproductive and Productive Performance of Dairy Animals Maintained at Mulitary Dairy Farm Lahore Cantt.
by Abrar Ahmed | Dr. Mian Abdul Sattar | Dr. Amjad Riaz | Prof. Dr. Muhammad.
Material type: ; Format:
print
Publisher: 2012Dissertation note: NILI RAVI BUFFALOES
Average age at maturitywas 829.11 ± 30.34 days. Heiferscalved in1999 and 2008 were found to have maximum and minimum age at maturity, respectively. Heifers calved during the winter and dry summer season were found to have maximum and minimum age at maturity, respectively. Mean birth weight of calves was found 28.16±2.8 kg. The effect of birth weight on age at maturity was found to be significant and it was observed that 1 kg increase in birth weight resulted in 16 days decrease in age at maturity. Average age at first conception was 923.37 ± 30.34 days. Heifers calved in 1996 and 2008 were found to have maximum and minimum age at first conception, respectively. Heifers calved during the winter and dry summer season were found to have maximum and minimum age at first conception, respectively. Average age at first calving was 1295.66 ± 62.91 days. Heifers calved in 2002 and 2008 were found to have maximum and minimum age at first calving, respectively. Heifers calved during the dry summer andautumn season were found to have maximum and minimum age at first calving, respectively. Average service period was 164.1 ± 30.15 days. Maximum and minimum service period was in year of 2002 and 2008, respectively. In dry summer and autumn season, service period was maximum and minimum, respectively. In 11th and 10thlactation, service period was maximum and minimum, respectively.Average services per conception were 1.95 ± 0.23. Maximum and minimum services per conception were in 2005 and 2008, respectively. In humid summer and autumn season, services per conception were maximum and minimum respectively. In 11th and 9th lactation, the services per conception were maximum and minimum, respectively. Average gestation period was 310.90 ± 3.07 days. Maximum and minimum gestation periodwas in 2000 and 2009 2009 respectively. In humid summer and dry summer season, gestation period?
was maximum and minimum respectively. In 12th and 8th lactation, the gestation period was maximum and minimum, respectively. Average calving interval was 474.59 ± 29.88 days. Maximum and minimum calving interval was in 2005 and 2001, respectively. In dry summer and autumn season, calving interval was maximum and minimum respectively. In 11th and 10th lactation, calving interval was maximum and minimum, respectively. Average milk yield was 1973.15 ± 53.23 liters. Maximum and minimum milk yield was in 2010 and 2002, respectively. In winter and spring season, milk yield was maximum and minimum respectively. In 4th and 11th lactation, milk yield was maximum and minimum, respectively. Average lactation length was 295.23 ± 64.30 days. Maximum and minimum lactation length was 2001 and 2000, respectively. In spring and humid summer season, lactation length was maximum and minimum respectively. In 10th and 11th lactation, lactation length was maximum and minimum, respectively. Average dry period was 184.93 ± 30.99 days. Maximum and minimum dry period was in 2005 and 2001, respectively. In dry summer and autumn season, dry period was maximum and minimum respectively. In 11th and 10th lactation, dry period was maximum and minimum, respectively.
CROSSBRED COWS
Average age at maturity was 828.33 ± 20.39 days. Heifers calved during the year of 1998 and 1997 were found to have maximum and minimum age at maturity, respectively. Heifers calved during the humid summer and winter season were found to have maximum and minimum age at maturity, respectively. In 15/16 and 1 /2 crossbred level, age at maturity was maximum and minimum, respectively. Mean birth weight of crossbred cow calves was found 23.35 ± 2.8 kg. The effect of birth weight on age at maturity was found to be significant and it was observed that 1 kg increase in birth weight resulted in 07 days decrease in age at maturity. Average age at first conception was 944.18 ± 30.42 days. Heifers calved in 2004 and 1996 were found to have maximum and minimum age at first conception, respectively. Heifers calved during the humid summer and dry summer season were found to have maximum and minimum age at first conception, respectively. In 15/16 and 1 /2 crossbred level, age at first conception was maximum and minimum, respectively. Average age at first calving was 1245.29 ± 40.96 days. Heifers calved in 2008 and 1995 were found to have maximum and minimum age at first calving, respectively. Heifers calved during autumn and winter season were found to have maximum and minimum age at first calving, respectively. In 7/8 and 5/8 crossbred level, age at first calving was maximum and minimum, respectively. Average service period was 252.83 ± 32.56 days. Maximum and minimum service period was in year of 2006 and 2010, respectively. In winter and humid summer season, service period was maximum and minimum respectively. In 10th and 8th lactation, service period was maximum and minimum, respectively. In 7/8 and 15/16 crossbred level, service period was maximum and minimum, respectively. Average services per conception were 2.13 ± 0.31. Maximum and minimum services per conception were in 1999 and 2002, respectively. In winter and dry summer season, services per conception were maximum and minimum respectively. In 9th and 8th lactation, the services per conception were maximum and minimum, respectively. In 15/16 and 5/8 crossbred level, services per conception were maximum and minimum, respectively. Average gestation period was 278.90 ± 4.36 days. Maximum and minimum gestation period was in 2010 and 2003 respectively. In autumn and winter season, gestation period was maximum and minimum, respectively. In 2nd and 7th lactation, the gestation period was maximum and minimum, respectively. In 7/8 and 15/16 crossbred level, gestation period was maximum and minimum, respectively. Average calving interval was 507.03 ± 33.61 days. Maximum and minimum calving interval was in 2006 and 2000, respectively. In winter and humid summer season, calving interval was maximum and minimum respectively. In 11th and 10th lactation, calving interval was maximum and minimum, respectively. In 5/8 and 15/16 crossbred level, calving interval was maximum and minimum, respectively. Average milk yield was 2141.05 ± 90.10 liters. Maximum and minimum milk yield was in 2008 and 2003, respectively. In dry summer and autumn season, milk yield was maximum and minimum respectively. In 11th and 1st lactation, milk yield was maximum and minimum, respectively. In 15/16 and 3/4 crossbred level, milk yield was maximum and minimum, respectively. Average lactation length was279.31 ± 8.33 days. Maximum and minimum lactation length was 2001 and 2003, respectively. In dry summer and autumn season, lactation length was maximum and minimum respectively. In 11th and 5th lactation, lactation length was maximum and minimum, respectively. In 3/4 and 15/16 crossbred level, lactation length was maximum and minimum, respectively. Average dry period was 238.64 ± 33.93 days. Maximum and minimum dry period was in 2006 and 2000, respectively. In winter and humid summer season, dry period was maximum and minimum respectively. In 5th and 11th lactation, dry period was maximum and minimum, respectively. In 5/8 and 15/16 crossbred level, dry period was maximum and minimum, respectively.
Availability: Items available for loan: UVAS Library [Call number: 1624,T] (1).
5.
Prevalence Of Brucellosis In Dairy Animals And Their Handlers In District Bannu, Khyber Pakhtoon Khwa
by Azmatullah Khan | Dr. Amjad Riaz | Dr. Hassan | Dr. Mian Abdul Sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1641,T] (1).
6.
Effect Of Biostimulation During Estrus Synchronization Protocols On Pregnancy Rate In Buffalo
by Abdul Nasir | Dr. Amjad Riaz | Dr. Aqeel | Dr. Mian Abdul Sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1666,T] (1).
7.
Effect of Timing of Artificial Insemination and Ovulation in Relation to Onset of Standing Heat on Pregnancy Rate in Nili-Ravi Buffaloes
by Umair Riaz | Prof. Dr. Nasim Ahmad | Dr. Mian Abdul Sattar | Prof. Dr.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Artificial insemination (AI) is one of the first and most influential biotechnologies of animal agriculture. To be beneficial artificial insemination must be done properly. There are many factors involved in quality of AI performance, including insemination technique, technician, semen quality, cleanliness, etc. However, the single most important factor in achieving high conception rate in buffalo is the timing of AI relative to the timing of ovulation. Application of AI is difficult because of poor estrus expression by buffaloes and poor estrus detection by owners, a highly variable duration of estrus and the difficulty to predict time of ovulation. The objective of the present study was to determine the effect of A.I at different time intervals (0, 12, 24 hours after the onset of standing heat) in relation to time of ovulation on pregnancy rate in buffalo. For this purpose, buffaloes with average body condition 2.5 ± 0.5 at Livestock Experimental Station Bhuneki, Buffalo Research Institute, Pattoki District Kasur during breeding season (September to December) were exposed to teaser bull two times in a day at 12 hours interval (6:00 am in morning and 6:00 pm in evening) to determine the standing heat (0h). Buffaloes, allocated randomly, at different time intervals i.e., either 0, 12 or 24 hours after the onset of standing heat were bred artificially using frozen thawed semen. Trans-rectal ultrasonography (HS 1500V; Honda; Japan, with 7.5 MHz transducer) was used to determine ovarian follicular development and ovulation, from onset of standing heat till ovulation after every 12 hours. Pregnancy status was determined by ultrasonography, 30 to 40 days post AI. Results revealed that the overall time of ovulation from onset of standing heat averaged 35.33 ± 0.21 h. Pregnancy rates were 26% (8/30) at 0 h, 37% (10/27) at 12 h and 53% (15/28) at 24 h after the onset of standing heat. It is concluded that timing of ovulation is about 35 h and 24
maximum pregnancy rate are achieved when bred after 24 hours after the onset of heat in buffaloes.
Availability: Items available for loan: UVAS Library [Call number: 1694,T] (1).
8.
Effect Of Control Internal Drug Release & Human Chorionic Gonadotropin Administration On Day 7 After Fixed Time
by Ali Husnain | Prof. Dr. Nasim Ahmad | Dr Mian Abdul sattar | Prof. Dr.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1697,T] (1).
9.
Association Between Numbers Of Ovarian Follicles And Fertility In Nili-Ravi Buffaloes
by Qaisar Shahzad | Dr. Amjad Riaz | Dr. Jawad Nazir | Dr. Mian Abdul Sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: 6.1: Back Ground:
The water buffalo (Bubalus bubalis) is an important animal of livestock species. It has an essential role in the economy of developing countries. Buffalo is being used as dairy, beef and draught purpose. Despite so many qualities, the reproductive potential of the buffalo is lower than cattle. One of the reasons for low reproductive potential is smaller ovaries and low number of ovarian follicles. Due to these reasons superovulation and embryo transfer has not been much successful in buffaloes. There is no tool through which reproductive performance of buffaloes can be phenotyped. Antral follicular count can be a tool on the basis of which reproductive performance of buffaloes can be phenotyped
6.2: Hypothesis:
Higher the antral follicular count higher will be the fertility in Nili-Ravi buffaloes.
6.3: Methodology:
Ten Nili-Ravi heifers were used in the study to measure repeatability of antral follicular. Each heifer was scanned on alternating days from day 1 of estrous cycles to day 9 of the successive estrous cycle. Antral follicles were counted in different follicular waves and repeatability of antral follicles was assessed. In the 2nd experiment of the study 10 heifers were used to count antral follicles from day 1 to day 10 of the estrous cycle, on the basis of antral follicular count animals were divided into three groups, Low (1-4), Intermediate (5-8) and High (?9).Blood sampling was done on 7th day of estrous cycle to measure progesterone concentration of 3 animals from each group and an association was developed between antral follicular count and progesterone concentration. In the 3rd experiment, 25 Nili-Ravi buffaloes were used to antral follicles from day 1 of estrous cycle to day 10 of estrous cycle. , on the basis of antral follicular count animals were divided into three groups, Low (1-4), Intermediate (5-8) and High (?9). In the next heat animals were artificially inseminated and were checked for pregnancy on 36th and 60th day by using ultrasound. Animals pregnant on 60th day were considered as pregnant. After that association was developed between antral follicular count and fertility.
6.4: Results:
Buffalo can be phenotyped on the basis of antral follicular count. Follicular counts are highly repeatable in different follicular waves of same estrous cycle (0.83) and different estrous cycles (0.85) within individual animals. Follicular counts are highly positively correlated (0.91) with progesterone concentration. Higher the antral follicular count, higher should be the progesterone concentration. Antral follicular count is highly positively associated with (0.99) with fertility. Higher the antral follicular count higher will be the fertility.
Availability: Items available for loan: UVAS Library [Call number: 1734,T] (1).
10.
Effect Of Cholesterol Addition On Cryopreservation Of Spermatozoa And Field Fertility Of Nili Ravi Buffalo
by Abdul Razzaq | Dr. Mian Abdul Sattar | Dr. Amjad Riaz | Dr. Muhammad Avais.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1821,T] (1).
11.
A Study On Superovuation Protocol For The Development Of Embryo Transfer Technique In Mice
by Muhammad Ameen jamal | Dr. Amjad riaz | Dr. Aamir | Prof. Dr Mian abdul sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1900,T] (1).
12.
Effect Of Butylated Hydroxytoluene (Bht) On Beetal Buck Semen Following Cryopreservation
by Abdul Rehman | Prof.Dr. Mian Abdul Sattar | Dr. Amjad Riaz | Dr. Muhammad Ijaz.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1977,T] (1).
13.
Effect Of Cholesterol Loaded Cyclodextrins (Clc) On Post-Thaw Quality In Canine Spermatozoa
by Junaid khan | Prof. Dr.Nasim ahmad | Dr. Asim khalid | Prof. Dr. Mian abdul sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2005,T] (1).
14.
Standardization Of Bovine Serum Albumin (Bsa) For Cryopreservation Of Beetal Buck Semen
by Muhammad Shahzad | Prof. Dr. Mian abdul sattar | Prof. Dr | Prof. Dr. Naseem ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2018,T] (1).
15.
Effect Of Cholesterol-Loaded Cyclodextrin (Clc) Addition On Egg Yolk Ratio In Semen Extender And Post-Thaw Quality Of Buffalo Bull Sperm
by Mehboob Ahmed | Dr. Mushtaq Ahmad | Prof. Dr. Mian Abdul Sattar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2085,T] (1).
16.
Optimization Of Strontium Chloride For Parthenogenetic Activation Of Mouse Oocytes
by Arslan Mahmood Ahmad (2007-VA-67) | Dr. Amjad Riaz | Dr. Aqeel Javeed | Prof. Dr. Mian Abdul Sattar.
Material type: ; Format:
print
Publisher: 2014Dissertation note: There are two main methods by which activation can be performed: (i) physical methods and (ii) chemical methods. Physical methods include electrical stimulation, temperate and mechanical ways, whereas the chemical methods comprise of different artificial chemical agents, including strontium chloride, calcium ionophores, ethanol that promote to rise in intracellular Ca2+ oscillations, cycloheximide, that inhibit protein synthesis and 6-DMAP (6-dimethyl amino purine) which inhibit protein phosphorylation.
The contribution of both maternal and paternal genomes is required for thedevelopment to full term of mammalian embryos. However, the percentage of parthenogeneticallyactivated embryos developing to blastocyst stage is lower as compared to normal fertilized embryos. (Renard et al. 1991).In mouse, strontium chloride has been successfully employed in manydifferent studies to induce artificial oocyte activation. The role of strontium to induce calcium oscillations appears to be more physiologically sound than alternativemethods of oocyte activation that produce a monotonic rise in calcium.Strontium chloride (SrCl2) is recognized as one of the most popular parthenogenetic agents for mouse oocytes activation and induces calcium oscillations leads to improved activation rate and blastocyst formation. (Locham-kaplan et al. 2003) (Satoshi et al. 2006).
The diploid parthenogenetic oocytes have more developmental competence as compared to haploid form(Liu et al. 2002). A substancecytochalasin B (CB) prevents the release of the second polar body after activation of mammalian oocyte which results in diploid form of embryo (Fukui et al. 1992) and it may also contribute to prevent fragmentation and degradation of embryos ( Yi and Park 2005).
Parthenogenetic oocyte activation technique is mainly used in cloning and is a key step for nuclear transfer for cloning. The technique is also useful for understanding of physiological mechanisms of fertilization and early embryonic development. Embryonic stem cells can be derived from fertilized embryos. The stem cells which are produced by parthenogenetic activation have the same totipotency and proliferation as formed by normal sperm-egg fertilization..( Ju et.al 2008). Resultantly, parthenogenetic activation technology has become a target of reproductive biology. This technology can also be used to establish embryonic stem cell lines (Mizutani et al. 2004) and embryonic stems cells are the fundamental source in field of regenerative medicine; used to treat many diseases such as diabetes, beta thalassemia, heart infarction etc by providing patient specific replacement cells.
Mouse is one of the most commonly animal models used for parthenogenetic activation. The other animals which have been used for parthenogenetic activation include rabbits, cattle, sheep, horses, monkeys and pigs. Parthenogenetic embryos are failed to develop to term, due to genomic imprinting, an epigenetic change of certain genes, depending on the parent of origin.(Uranga and Arechaga 1997). The studies pertaining to parthenogenetic activation technology for mouse oocytes is extremely limited at present (Mizutani et al. 2004).
Availability: Items available for loan: UVAS Library [Call number: 2188,T] (1).
17.
Effect of Estradiol Benzoate (EB) and Equine Chorionic Gonadotropin (eCG) on Ovarian Response and Conception Rate in Control Internal Drug Release (CIDR) Based Estrus Synchronization Protocol in Nili Ravi Buffalo
by Abdul Saboor Khan (2008-VA-116) | Prof. Dr.Nasim Ahmad | Prof.Dr. Mian Abdul Sattar | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Blank CD. Availability: Items available for loan: UVAS Library [Call number: 2239-T] (1).
18.
Comparative Effect Of Cidr Based Estrus Synchronization Protocol With Or Without Gnrh In Non-Descript Cows During Low Breeding Season
by Muhammad Bilal (2008-VA-152) | Prof. Dr. Mian Abdul Sattar | Dr. Muhammad Anwar | Dr. Muhammad Rizwan Yousuf | Dr. Muhammad Lateef.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The livestock sector occupies a peculiar position in the national plan of economic development in Pakistan. It subscribed approximately 55.9 percent to the agricultural value added and 11.8 percent to national GDP with a growth rate of 2.7 % in 2013-14 (Anonymous 2014). Dairying has become an important subsidiary source of income for thousands of rural families with an important role in generating earning opportunity. In Pakistan dairy sector is developing and commercializing at a rapid pace to meet increasing requirement of milk and other dairy products (Dongre et al. 2011).
Pakistan owns renowned breeds of dairy buffaloes (Nili-Ravi and Kundi) and cattle (Sahiwal, Red Sindhi, Thari and many others). Cattle in Pakistan belong to genus Bos indicus. According to the latest livestock census (Anonymous 2006), out of 29.6 million cattle, 46% (13.6 million) have been described as non-descript. Non-descript cattle do not fall in any defined breeds of cattle. Milk production of non-descript cows is < 1000 lit per lactation in mountains area of NWEP pakistan (Khan and Usmani 2005). As non-descript breeds make up the largest group of cattle in Pakistan, there is a dire need to work on genetic improvement of these animals. Artificial insemination, the best tool for genetic up-gradation in dairy cattle is applied only in 11.1% cows in Pakistan (Anonymous 2006). The main hindrances are small sized scattered herds and lack of experienced technical manpower in the field. Estrus synchronization of a large number of animals and timed insemination can be used to overcome these hindrances .The technique may also help in
11
reducing a prolonged calving interval and postpartum anestrus and seasonality of breeding in these animals (Zafar et al. 2008).
Estrus synchronization widely practiced in temperate dairy cattle in developed countries (Hansen and Arechiga 1999). Before launching a large scale estrus synchronization program in non-descript cattle, there is a need to assess the efficacy of various synchronization protocols in terms of estrus incidence, intensity and conception rate. Additionaly, distinct differences have also been reported between Bos taurus and Bos indicus in terms of estrus duration and intensity of expression of estrus sign (Mattoni and Ouedraogo 2000). The low estrus intensity and less duration of estrus signs of Bos Indicus are due to smaller diameter of follicle as compared to that of Bos Taurus (Bo et al. 2003).
Developing successful methods for synchronizing estrus and ovulation in cattle has been a major research interest. Ultimately, the goal has been to achieve precise synchronization of ovulation so that cattle can be inseminated without regard to estrus detection. One method to increase conception rates is to use hormonal treatments in zebu breeds for synchronizing ovulation and for timed artificial insemination (TAI). Hormonal programs for synchronizing ovulation to control the follicular and luteal phases and estrus behavior have been used in Bos taurus cows and heifers(Castellanos et al. 1997), and Bos Indicus cows (Pinheiro et al. 1998).
The intensity and duration of estrus behaviors during the estrous cycle is highly variable among individuals. More commonly animals are diagnosed to be in estrus based on the mounting or standing to be mounted, appearance of mucus discharge, and other physical activities (Van Eerdenburg et al. 2002). Scoring system were established on the basis of observed estrus signs and most of them have taken mounting and standing to be
12
mounted behavior as the most reliable signs to predict ovulation time in Bos taurus cows (Roelofs et al. 2005). However, it is now well acknowledged that the expression of estrus behavior change with breed of cows (Naidu and Babu Rao 2006).
Fertility is an important parameter to assess the efficacy of estrus synchronization. Bos indicus cows after treatment with CIDR conception rate in adult cows 40% and in heifers 20% (Singh et al. 2006). CIDR may have ability to overcome the problems in field condition and increasing the reproductive efficiency by minimize the hindrance anestrus postpartum cows. In Pakistan research work have been done to evaluate the efficacy of CIDR for conception rate in indigenous cattle. Until now no study conducted on non- descript cattle. Therefore, it is hypothesized that CIDR+GnRH can provide better in vivo fertility compared to CIDR. Availability: Items available for loan: UVAS Library [Call number: 2248-T] (1).
19.
Comparative Effect Of Alpha Lipoic Acid And Butylated Hydroxytoulene On Post Thaw Quality Of Buck Semen
by Muhammad Khurram Shahzad | Prof. Dr. Mian Abdul Sattar | Dr. Mushtaq Ahmad | Dr. Muhammad Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Amongst different livestock species, goats and sheep are the major source of livelihood for over a million livestock farmers in Pakistan. Total goat population in Pakistan is estimated to be 66.6 million. These animals are mostly kept by small holders for whom these are only source of their livelihood. Milk production from goats is 0.822 million tonnes while mutton production from both sheep and goats is 0.657 million tonnes (Anonymous 2014). Pakistani people mostly prefer the goat meat over sheep.
All irrigated areas of Punjab including district Faisalabad, Sahiwal, Sargodha, Jhang, Jhelum, Lahore and Multan are the habitat of Makhi Cheeni Beetal goats. The color of its body coat is red spotted or golden brown with white patches. Its body is very well developed and compact. Males have long spiraled horns while females have shorter. It has roman nose with pendulous broad and long ears. It has long teats and well developed udder. Female weighs 37kg and males 46kg. Twins or triplets births are more than 50%. In 130 days of lactation period, there is 290 liters milk yield (Shah et al. 2001).
Some breeds of goats especially dairy goats have more demand than the others and these bucks are not available everywhere. To cope with this situation artificial insemination techniques is necessary. Artificial insemination plays a great role in increasing the economics by spreading the superior genetics within a short period of time.
Semen is processed by different methods but cryopreservation is considered to be the best method. Cryopreservation has been reported to compromise the quality of processed semen resulting in the loss of sperm motility, viability, in-vivo fertilizing ability, deterioration of plasma membrane and acrosomal integrity, apoptosis and damage of deoxyribonucleic acid (DNA) (Medeiros et al. 2002; Purdy 2006a). Sperm damage may occur due to various factors like osmotic stress, oxidative stress, low-temperature exposure and combination of different factors (Sarıözkan et al. 2009). Thawing of semen may also cause osmotic changes and the sperm quality is further decreased. It is generally accepted that sperm viability is reduced by as much as 50% during the process of semen cryopreservation (Watson, 2000).
Extension of buck semen with egg yolk containing extender may be more injurious to sperms. This is due to the presence of coagulating enzymes of bulbourethral origin named as egg yolk coagulating enzymes (EYCE). EYCE decreases the tenacity of chilled or frozen semen (Roy, 1957). EYCE also catalyze the conversion of egg yolk lecithin into lsolecithin and fatty acid, thus sperm membrane become more fusogenic due to hydrolysis. So there is increase in chromatin decondensation and acrosomal reaction that is harmful for sperm (Leboeuf et al. 2000). Due to excess of poly unsaturated fatty acids (PUFA) in sperms, they are more susceptible to lipid peroxidation (Cassani et al. 2005). Lipid peroxidation of PUFA lead to production of reactive oxygen species (ROS) (Alvarez et al. 1995). Small amount of ROS are normally involved in capacitation, acrosmal reaction and ultimately fertilizing ability of sperms. But when the ROS are produced in excess
Introduction
3
amount, these may compromise the enzymatic function and sperm fertility (Baumber et al. 2000). At 4-5 ºC the motility and plasma membrane integrity is decrease with the passage of time which ultimately leads to decrease in fertility. One of the cause of this decrease is production of ROS by the lipid peroxidation of spermatozoa’s membrane (Storey et al. 1998). Major decrease in sperm motility and fertility occur during phase transition from liquid crystalline to gel phase (Chakrabarty et al. 2007). Lipid peroxidation leads to irreversible loss in motility and damage to DNA of sperm (Maxwell et al. 1996). Motility of sperm is adversely effected with ROS, when the ROS harm the plasma membrane and acrosomal integrity which ultimately leads to fragmentation of DNA. Sperms have their own antioxidants system which include the glutathione (GSH) , GSH peroxides, superoxide dismutase, catalase and chelators of transferrin, lactoferrin and ceruplasmin (Agarwal et al. 2002). Normally the ROS production and scavenging are in equilibrium but during the semen preservation the excessive production of ROS (superoxide, hydroxyl, hydrogen peroxide, nitric oxide, peroxynitrile) with low level of scavenging system and antioxidants leads to oxidative stress. During the process of freezing and thawing the natural antioxidants systems are unable to stop lipid peroxidation. Therefore a powerful antioxidant system should be used to avoid the cryo-injuries and lipid peroxidation (Irvine 1996).
Different antioxidants are being used i.e. fetuin (F), amino acid (AS), cysteine (CY) taurine, glutathione (GSH) glutathione peroxidase (GSH-PX), catalase (CAT), superoxide dismutase (SOD) glutamine, hyaluronan, trehalose, Alpha lipoic acid (ALA) and Butylated Hydroxytoulene (BHT) (Atessahin et al. 2008; Bucak et al. 2009; Taşdemir et al. 2014). Addition of antioxidants to semen extenders are considered to improve the quality of semen (Rao et al. 2013). ALA is a short chain fatty acid which act as an antioxidant in both aqueous and lipid environments, its therapeutic effects in other tissues like brain (Piotrowski et al. 2001), heart, kidneys and testicles has already been
Introduction
4
discussed. It is called as universal antioxidant because of its effect in different parts of body. It is not only involve in scavenging the ROS but also activate the body antioxidants systems against ROS. ALA reduced to dithiol form called dihydrolipoic acid (DHLA) which is an excellent antioxidant (Handelman et al. 1994). ALA also regenerates vitamin C from reduced vitamin C in the presence of glutathione (GSH) which also enhance the antioxidant activity (Ibrahim et al. 2008). BHT, a phenolic lipophilic antioxidant that has antiviral activity, have the ability to relieve the cold shock in spermatozoa from several animal species. It stops the auto oxidation by converting the peroxide radical to hydroperoxide as it is also called as synthetic analogues of Vit E (Memon et al. 2011). BHT acts as a membrane lipid protectant which reduces the changes in permeability of sperm plasma membrane in cold shock (Graham et al. 1992). BHT minimizes the effect of cold shock on semen (Shoae et al. 2008), boar (Roca et al. 2004) and goat (Khalifa et al. 2008). Availability: Items available for loan: UVAS Library [Call number: 2254-T] (1).
20.
Comparison Between Aspiration And Slicing Methods For Retrieval Of Oocytes In Bovine
by Muhammad Husnain (2008-VA-281) | Prof. Dr. Mian Abdul Sattar | Dr. Qaiser Shahzad | Dr. Amjad Riaz | Dr. Muhammad Avais.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Livestock contribution to agriculture stood at 55.9 percent while it contributes 11.8 percent to the national GDP during 2013-14. Buffalo, cattle, sheep and goat population in Pakistan is 34.6, 39.7, 29.1 and 66.6 million numbers during 2013-14. Total milk production from buffalo and cattle as major milk producing animals is 31,252 and 18,027 (000 tons) (Economic Survey of Pakistan 2013-14).
Advanced biotechnologies coming from different areas of biological sciences exhibit great promise to enhance the efficiency of livestock production. From these technologies one such biotechnology is the use of in vitro maturation of follicular oocytes and in vitro fertilization for production of livestock embryos in laboratory. Proper oocytes recovery and their selection in the laboratory are of great importance for successful in vitro embryo production. Total one hundred and forty four ovaries (n=144) from cattle (72 ovaries) and buffalo (72 ovaries) were collected and 223 oocytes were retrieved from these ovaries. Average oocytes per ovary were 1.66 + 0.43 oocytes per ovary were obtained via aspiration and 1.89 + 0.00 average oocytes per ovary through slicing method from cattle ovaries. Average 1.55 ± 0.55 oocytes per ovary via aspiration and 1.53 ± 0.20 oocytes per ovary through slicing from buffalo ovaries.
Overall grade-A oocytes were 28 (40) percent with aspiration in cattle and 25(36.76) through slicing method. In buffalo overall grade-A oocytes retrieval was obtained in percentage as 20 (44.44) and 26 (52) through aspiration and slicing methods respectively. Grade-B oocytes recovery obtained was in percentage as 23 (33.82) with slicing and 19 (31.67) through aspiration technique from cattle ovaries.
Summary
26
Commonly used methods of recovery of oocytes from slaughterhouse animals are aspiration and slicing. Recovery rate of oocytes is different from slaughterhouse ovaries. Aspiration is the best method for retrieval of good quality oocytes from slaughterhouse bovine ovaries because it gave more good quality oocytes in less time than slicing method. In this study, it is found that weight of ovary and no. of follicles/ovary in cattle have strong correlation of 71% existed between weight of ovary and no. of follicles /ovary in buffalo was observed. Correlation between average number of follicles on ovary and weight /ovary was stronger in cattle. The more the number of follicles present on the ovaries and more weight of the ovary, the more will be the recovery of oocytes.
In cattle average number of follicles was 10.09 ± 0.30 and when it was checked in buffalo, differed significantly and it was found as 7.16 ± 0.19 on an average per ovary. Likewise weight of buffalo in this study was differed significantly from cattle 4.04 ± 0.10 and 7.62 ± 0.15 respectively. It is suggested that oocytes retrieval should be done in buffalo using aspiration method to retrieve better quality oocytes.
It is concluded that aspiration is the suitable method for retrieval of good quality oocytes from slaughterhouse buffalo ovaries because it gave more good quality oocytes in less time than slicing method. But both methods have minor difference between recovery rates but aspiration is more convenient than slicing and it yields more quality oocytes. It is also found that there is very strong correlation existed between average weight of ovary and number of follicles per ovary and the both parameters play a great help for more quality and quantity oocytes. Availability: Items available for loan: UVAS Library [Call number: 2279-T] (1).
(1 votes)
21.
Effect Of L-Cysteine And Glutathione On Post Thaw Quality Of Sahiwal Bull Spermatozoa
by Farhan Younas (2007-VA-495) | Prof. Dr. Mian Abdul Sattar | Dr. Syed Murtaza Hasan Andrabi | Prof. Dr. Nasim Ahmad | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Freezing and thawing of semen leads to production of reactive oxygen species (ROS)
due to plasma membrane lipid peroxidation. Because of this semen quality can be
compromised. To overcome this problem, antioxidants have been used in cryopreservation
medium. Glutathione and cysteine have thiol groups which penetrate into the cell and protect
it from oxidative stress. In this study, effect of different concentrations of cysteine and
glutathione on post thaw quality of Sahiwal bull spermatozoa was determined.
Semen was collected with artificial vagina from five mature regular donor Sahiwal
bulls kept at the Semen Production Unit Qadirabad, Sahiwal. Semen samples possessing
>60% motility and >500x10
6
sperm/ml were included in study. After collection, semen
samples from five bulls were pooled, divided into seven equal aliquots and kept at 37 ºC in
water bath. After that dilution was done with Tris citric egg yolk extender having different
concentrations of cysteine and glutathione as Con (0.0 mM), C1 (1.0 mM cystein), G1 (1.0
mM glutathione), CG0.5/1(0.5 mM Cysteine+1.0 mM glutathione), CG1/0.5 (1.0 mM
cysteine+0.5 mM Glutathione), CG0.5/0.5 (0.5 mM cysteine+0.5 mM glutathione) and
CG1/1 (1.0 mM cysteine+1.0 mM glutathione). Diluted samples were cooled to 4ºC in two
hours and equilibrated for 4 hours at 4
o
C. After that they were packaged into 0.5 ml French
semen straws (20x10
6
sperm/straw). All semen straws were placed 4cm above liquid nitrogen
surface in vapors for 10 minutes. Then, semen straws were plunged into liquid nitrogen for
freezing and stored until post thaw analysis. The experiment was repeated for five times
(replicates = 5). Four semen straws/treatment were thawed for 30 seconds in water bath at
37ºC and evaluated for visual motility, plasma membrane integrity (PMI), acrosome integrity,
mitochondrial trans membrane potential and CASA motility parameters and kinematics.
42
Summary
PMI in group CG0.5/0.5 was significantly higher (40.00±1.42 %) as compared to Con
26.67±0.80 (P<0.5). Plasma membrane integrity in groups CG1/1, CG0.5/1, G1 and C1 was
significantly higher (36.00±1.88 %, 36.20±1.07 %, 33.60±1.21 % and 32.80±0.80 %
respectively) as compared to Con (26.67±0.80 %) (P<0.05). There was no significant
difference in C1 (32.80±0.80 %) and G1 (33.60±1.21 %) (P>0.05). In case of acrosome
integrity, NAR value of group CG0.5/0.5 was significantly higher (71.40±1.08 %) as
compared to Con (59.67±0.37 %) (P<0.05). All other groups also showed significant
differences as compared to Con (P<0.05). CG0.5/0.5 also showed significantly higher NAR
value (71.40±1.08 %) as compared to C1 (64.40±1.40 %) and G1 (67.60±2.07 %) (P<0.05).
CG0.5/0.5 had significantly higher value (71.40±1.08 %) as compared to CG1/0.5 and CG1/1
(65.60±0.81 % and 68.80±0.97 % respectively) (P<0.05). CG0.5/0.5 had significantly higher
subjective motility (54.00±1.88) as compared to Con (36.66±0.92)
Mitochondrial transmembrane potential of CG0.5/0.5 was significantly higher
(37.00±0.71 %) as compared to Con (25.33±1.28 %) (P<0.05). All the other treatment groups
also had higher mitochondrial transmembrane potential as compared to Con (P<0.05). In
groups of combination of cysteine and glutathione, CG0.5/0.5 showed significant difference
(37.00±0.71 %) as compared to CG1/1 and CG1/0.5 (29.00±1.00 % and 33.80±0.86 %)
respectively (P<0.05).
CASA results showed that CG1/1 had significantly higher motility as compared to the
control. But the percentage of progressive spermatozoa was significantly higher in
CG0.5/0.5. VSL of group CG0.5/0.5 was significantly higher (53.33±2.90 %) as compared to
Con (45.10±0.50 %). However, VSL, VCL, ALH and BCF did not vary significantly among
groups. STR and LIN of group CG0.5/0.5 were significantly higher as compared to the
control group.
43
Summary
In conclusion, addition of cysteine and glutathione in tris citric egg yolk extender
improved the post thaw quality of Sahiwal bull spermatozoa. In case of additive effect of
cysteine and glutathione, CG0.5/0.5 showed higher plasma membrane integrity, acrosome
integrity, mitochondrial transmembrane potential, progressive and rapid spermatozoa as
compared to CG0.5/1, CG1/0.5 and CG1/1.
44 Availability: Items available for loan: UVAS Library [Call number: 2318-T] (1).
22.
Effect Of Bio-Stimulation On Estrus Expression And Pregnancy Rate In Cidr Based Synchronization Protocol In Nili-Ravi Buffalo
by Abdul Waheed (2009-VA-133) | Dr. Aijaz ali Channa | Dr. Syed Murtaza Hassan Andrabi | Prof. Dr. Mian Abdul Sattar | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Our water buffalo (Bubalus bubalis) has much potential for production of milk. But this
animal has some problems regarding reproduction including delayed puberty, poor estrus behavior,
silent heat, long postpartum period and low conception rate by artificial insemination. This leads
to poor reproduction and hence great economic loss. Therefore, the requirement is to address these
problems efficiently and formulate more effective techniques for improvements. Researchers have
devised many estrus synchronization protocols (PGF2α, P4, GnRH, eCG, hCG etc.) that help
bringing many animals in heat and hence improve the reproductive performance when fixed time
artificial insemination is combined with them. But these protocols give inconsistent results when
they are applied on buffaloes making it necessary to improve the techniques.
This study was planned on the hypothesis that presence of bull (bio-stimulation), at the
time of synchronization, may play an important role in enhancement of estrus intensity and fertility
rate in Nili-Ravi buffaloes. Seventy one adult buffaloes were randomly selected from different
areas of field conditions and LRS (NARC) and subjected to CIDR based heat synchronization in
combination of either bio-stimulation or non-stimulation. The animals were observed for
behavioral estrus signs twice a day starting after 12 hours of CIDR removal till 96 hours.
Pregnancy diagnosis was done by rectal palpation 60 days post CIDR removal. Estrus response
and pregnancy rate were analyzed by Chi-square test using MINITAB version 15. Estrus signs and
total estrus intensity were compared by Mann Whitney U test. Difference was considered
significant at probability level of (P < 0.05).
In peri-urban areas, more animals from bio-stimulated group showed better behavioral
estrus signs, more total intensity score and significantly higher pregnancy rate as compared to nonSUMMARY
63
stimulated group of animals. At LRS (NARC), more animals from non-stimulated group were
found in behavioral estrus but intensity of heat signs was high in bio-stimulated animals.
Pregnancy rate was also higher in non-stimulated animals but the difference was not significant.
Overall, in this study, we got higher pregnancy rate in bio-stimulated animals than non-stimulated
group which indicates a positive response of bull stimulation on reproductive performance of Nili-
Ravi buffaloes who were synchronized with CIDR based estrus synchronization protocol. Availability: Items available for loan: UVAS Library [Call number: 2469-T] (1).
23.
Comparison Of Commercial Triladyl Extender With A Tris-Citric-Egg-Yolk (TCEY) Extender On Post-Thaw Semen Quality Of Nili Ravi Buffalo
by Muhammad Asad Ullah Khan | Prof. Dr. Mian Abdul Sattar | Prof. Dr. Nasim Ahmad | Prof. Dr. Mansur ud Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Cryopreservation of semen is the most important step for its usage in artificial insemination. Freezing of semen leads to a remarkable reduction in post-thaw semen quality. Therefore, selection of a better semen extender has always been considered priority that could serve as a good cryoprotectant.. Our semen production units (SPUs) have been using Tris based egg yolk semen extender since long time. Some modern SPUs like CEBG are using commercially available semen extenders for better post-thaw semen quality.
After collection pooled semen divided into two equal aliquots in separate sterilized test tubes and kept in water bath at 37 ºC. Semen was diluted with each of extender (TCEY and Triladyl) on the basis of sperm concentration (40x106sperm/ ml). Diluted semen was placed bottles and placed in safety cabinet cooled to 4 ºC over and equilibrated for 4 hrs. After equilibration semen was filled in 0.5 ml French straws (20x106sperm/ 0.5 ml). All semen straws placed in automatic freezer 4cm above liquid nitrogen surface in vapors for 10 minutes. Liquid Nitrogen vapors used in automatic programmable freezer to reduce temperature from 4 ºC to -180 ºC and then plunged into liquid nitrogen -196 ºC for freezing and was stored until analyzed. The experiment was repeated for seven times (replicates = 07)
CASA sperm motility parameter and kinematics were analyzed at Center of Excellence for Bovine Genetics (CEBG) Renala khurd District Okara. For further analysis frozen semen straws were brought to the Department of Theriogenology UVAS, Lahore. Effects of Triladyl and TCEY on post-thaw semen quality of the Nili Ravi buffalo semen were compared.
Summary
54
In Triladyl group, significantly (P<0.05) higher post-thaw motility (PTM %), Plasma membrane integrity (PMI, %),) DNA integrity (%), Live percentage was found. However, no significant (P<0.05) difference was found regarding NAR results between both groups. Sperm abnormalities were found significantly lower in Triladyl group as compared to TCEY group.
In overall assessment regarding and post-thaw CASA motility parameters, CASA motility, (PROG %), rapid (RAP %), medium (MED%), and slow (Slow, %) and sperm motility kinematics (VAP μm/sec), (VSL μm/sec), (VCL μm/sec), (ALH μm), (BCF HZ), (STR%) and (LIN%) Triladyl was found better than TCEY.
This was concluded that use of commercial semen extender Triladyl resulted in significantly better post-thaw semen quality as compared to Tris citric egg yolk (TCEY) extender. Availability: Items available for loan: UVAS Library [Call number: 2581-T] (1).
24.
Comparison Of Estradiol Benzoate And Gnrh In Cidr Based Superovulation Protocols For The Initiation Of Follicular Wave Emergence In Exotic And Crossbred Cattle
by Khalid Mahmood (2005-VA-114) | Dr. Amjad Riaz | Prof. Dr. Mian Abdul Sattar | Dr. M. Hassan Saleem.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Background: Livestock is a major contributor to the national (11.9%) and agriculture (55.4%)
economy in Pakistan. Milk and meat are major livestock products of Pakistan, which is ranked
fourth largest milk-producing country in the world. The growth rate of dairy sector is growing
very fast for last several years, however, the genetic potential of elite cows is continuously
deteriorating due to unavailability of reproductive biotechnologies such as embryo transfer.
Normally one calf per year can be obtained from elite mothers. Maximum number of offspring
can be obtained by superovulation. This will helpful in production of genetically superior
offspring in limited time thus resulting in maximum exploitation of genetic potential of elite cows.
In Pakistan more than 80% farmers are small holder having 2-3 animals with low genetic
potential. The use of elite mothers by these farmers is limiting due to high cost. Superovulation
is a strategy that can be used to make low cost embryos available for small holders. This will
result in maximum spread of genetic potential of superior females. Use of follicular wave
emergence based super stimulation and timed ovulation with help of CIDR can improve the
results of super ovulation protocols and may be an effective tool to improve the per unit time
embryo production.
Hypothesis: Use of EB or GnRH in CIDR based superovulation protocols may result in
improved super ovulatory response in cattle.
Methodology: This study was conducted at Centre of Excellence for Bovine Genetics Embryo
Transfer Wing Okara. Seventy Donor cows (mix of crossbred and Holstein Frisian) were selected.
Animals coming into natural heat, were randomly assigned into one of the following
superovulation protocols; (A) In first group (n=37), which was considered as control, on 8th day
after heat animals were palpated for presence of a good quality CL and super ovulatory treatment
CHAPTER 6
SUMMARY
38
i.e. twice daily FSH injection were started on “Day 11” of its cycle for four consecutive days. On
day 3rd of FSH treatment PGF2α was injected both in the morning and evening. Animals were
inseminated in the morning and evening on the 5th day of superovulation treatment and next day
morning based on detected heat. Embryos were collected from the animal on 7th day after first
insemination. A PG injection was administered to the animal three days after embryo collection.
(B) In second superovulation protocol (n= 15), the animals were palpated for the presence of a
good quality CL on “Day 8” and a CIDR was placed after the confirmation of CL. An injection
of 2mg EB (Estradiol Benzoate) was also administered on the same day. Super ovulatory
treatment was initiated by “Day 11” of its heat cycle as narrated in first superovulation protocol
with only difference of CIDR removal along with 7th dose of FSH. Animals were inseminated
with a single straw of semen if on heat in the next day morning (Day 16 after natural heat) or with
double straw at “3 pm” if not on heat in the morning. Animals were also inseminated on next day
morning if heat sustained till next day morning. Animal were collected by non-surgical flushing
seven days after first insemination at super estrus. A PG injection was also given to the animals
three days after embryo collection. (C) In third superovulation protocol (n=18), the animal were
palpated for the presence of a good quality CL on “Day 8” and a CIDR was placed after the
confirmation of CL. An injection of “2ml Dalmaralin” was administered on the same day. Super
ovulatory treatment was initiated by “Day 11” of its heat cycle in similar sequence as narrated in
second superovulation protocol. At least 5 animals of each category (i.e. Crossbred and Holstein
Frisian) were treated with each super stimulatory protocol. In conclusion, CIDR plus GnRH or
CIDR plus EB protocols are better than normal superovulation protocol for embryo production in
cattle.
Summary
39
Outcomes: This study remained helpful to improve the existing superovulation protocols for
bovines with promising results which will help the genetic improvement programs of bovine in
Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2636-T] (1).
25.
Comparison Of The Cryoprotective Effect Ofquailand Chicken Egg Yolk On The Freezability Of Nili-Ravi Buffalo Bull Spermatozoa
by Kamran Khaliq (2014-VA-812) | Prof. Dr. Nasim Ahmad | Prof. Dr. Mian Abdul Sattar | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pakistan is one of the most important agricultural country. Livestock as a subsector of agriculture contributes 11.4% in the national economy and 56.3% in agriculture. Buffalo is honored as the black gold of Pakistan and Nili-Ravi is the most important animal in this regard. Approximately 60% of the total milk produced within a country comes from buffalo. Artificial insemination is considered to be the most important tool for the prompt genetic improvement of livestock. Mammalian spermatozoa undergo many structural and biochemical changes during cryopreservation which may leads to an impaired fertility. Buffalo bull spermatozoa are sensitive and more prone to damage than cattle. Various experiments have been conducted in order to improve the post thaw semen quality which includes supplementation of egg yolk from different bird species that tends to work as a cryoprotectent against cold shock. In the present study it is assumed that replacement of chicken egg yolk with quail egg yolk in cryodiluents improves the freezability of Nili-Ravi buffalo bull spermatozoa.
Three adult Nili-Ravi buffalo bulls without any clinical reproductive anomaly and kept at Semen Production Unit (SPU), Qadirabad District Sahiwal were used in this study. Semen from these three experimental bulls was collected twice a day twice a week by using an artificial vagina already maintained at 42 ºC. Each bull was subjected to a minimum of seven replicas. Both semen samples were collected on each collection day from each of the three experimental bulls with an interval of about ten minutes between the ejaculates. After initial evaluation for sperm motility and concentration, both ejaculates collected from the same bull at the same collection day were pooled and again evaluated for sperm cell concentration. Every pooled semen sample was then divided into five aliquots and extended with one of the five experimental extenders namely A, B, C, D, and E in order to maintain a final concentration of 40 million spermatozoa per 0.54 ml of diluted semen. Instantly after dilution motility percentile for each semen sample was recorded. After filling, open ends of straws were sealed with polyvinyl pyrolidine powder and allowed to cool and then it was stored at 4-5 ºC for equilibration. Finally the samples were frozen and stored in liquid nitrogen. On post thaw evaluation, motility, live/dead count, plasma membrane integrity, acrosomal integrity, and DNA integrity along with CASA evaluation parameters for motility characteristics were performed. Data were analyzed by means of two way ANOVAand comparison between means was done by using Duncan’s multiple range test. Results showed that post-extension motility and DNA integrity did not differ significantly between extender A and E. While post-thaw motility, plasmolemma intactness, acrosomal integrity, and viable sperm count was found to be higher in extender E compared to extender A. Similarly, CASA evaluation factors like progressive sperm cells motility, straight line velocity, straightness, linearity index, and the percentile of rapidly moving spermatozoa were also significantly different among extender E and A. While curvilinear velocity, and amplitude of lateral head displacement was higher in extender B that contained 5% QEY. So it was concluded that substitution of 20% CEY with 20% QEY in cryodiluents resulted in an improved post-thaw motility, viable sperm count, plasmolemma intactness, and sperm kinematics. Furthermore, it was seen that reduction in the concentration of QEY from 20% to 5% resulted in decreased sperm motility characteristics.
Availability: Items available for loan: UVAS Library [Call number: 2686-T] (1).
26.
Effect Of Royal Jelly On Post Thaw Semen Quality Parameters Of Beetal Buck
by Muhammad Kaleem (2009-VA-190) | Dr. Abdul Rehman | Prof. Dr. Mian Abdul Sattar | Dr. Muhammad Avais.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pakistan is an agricultural country. Livestock plays a major role in the agriculture. Among livestock goat population is highest. In Pakistan Beetle is the most important breed of goat and is known as poor man’s cow. This goat is kept for dual purpose for milk and meat production. Artificial insemination is most valuable technique to improve the production and genetic potential of the goat. Cryopreserved semen has many biochemical, structural and functional problems. These detrimental effects produced by cryopreservation compromise the fertility of goat by decreasing post thaw motility, concentration, viability, plasma membrane and DNA integrity. These detrimental effects are due to many reasons among them production of reactive oxygen species is the most important ones. For last many years’ various solutions have been made to overcome the detrimental effect of reactive oxygen species. Various antioxidants are added in all domestic animals and their positive results have been demonstrated by many researchers. RJ is among these antioxidants which improve the sperm parameters.
RJ administration in the semen extenders has been shown to improve the post thaw motility, viability, DNA and plasma membrane integrity (PMI) of Beetal buck semen.
The study was conducted at Al-Haiwan Sires district Sahiwal, Punjab, Pakistan. For this study 3 regular semen donors Beetal bucks were used for semen collection. Semen was collected twice a week and total 7 collections was taken from each buck. After each collection semen of bucks was pooled to avoid individual buck variations and was divided into 5 equal parts in test tube containing extender with different concentration of royal jelly. These concentrations were . 0%, 0.5%, 1%, 1.5% and 2% RJ. Semen was cryopreserved in LN2.. On post thaw motility, livability, plasma membrane, DNA and acrosome integrity was evaluated.
Summary
27
Different royal jelly concentrations were analyzed by repeated measures analysis of variance (ANOVA).Duncan Multiple Range (DMR) test was used to compared the significant differences. Results of different group were expressed as mean ±SEM.
The results show that motility%, Plasma Membrane Integrity %, viability %, Normal Apical Ridge % and DNA integrity% of sperms was significantly high at 1% royal jelly concentration as compare to control and other treatment group. Availability: Items available for loan: UVAS Library [Call number: 2722-T] (1).
27.
Determination Of Comparative Effect Of Two Eeg Yolk Based Extenders On Post Thaw Semen Quality Of Sahiwal Bull
by Shahid Ali (2010-VA-05) | Prof. Dr. Main Abdul Sattar | Prof. Dr. manzoor Ahamd | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Cryopreservation of semen is the principal step for its usage in artificial insemination. Freezing of semen leads to reduction in post thaw semen quality. Glycerol has the cryoprotective properties that led to preserve spermatozoa. Egg yolk is also a basic constituent of semen extenders. Beneficial effect of egg yolk on sperm cryopreservation is plasma membrane protector. Tris based extenders are commonly used for semen cryopreservation of bulls, rams and bucks. Based on the economics and beneficial effects of extenders on bovine post thaw semen quality, tris based commercial as well as custom made semen extenders requiring egg yolk addition, needs to be considered for further studies. This study had been designed to determine the comparative effect of two egg yolk based extenders on post-thaw semen quality (Triladyl™and TCEY) on Sahiwal bulls. Semen was collected using an artificial vagina having temperature of 42 ºC from five adult Sahiwal regular donor bulls, raised at Center of Excellenc for Bovine Genetics (CEBG) Renalakhurd, Okara.Seven replicates of the experiment were performed. Volume, concentration and motility of ejaculates was evaluated. Semen samples having motility >60% and sperm/ml >500x106 were included in study. After evaluation, semen samples were pooled, divided into two aliquots of equal volume and kept in water bath at 37ºC. One aliquot was extended with tris citric acid egg yolk extender (TCEY) and other was extended with commercially available extender (Triladyl™). Pre-freeze CASA sperm motility parameters and kinematics of these extended aliquots were assessedat CEBG. After that, extended semen was cooled to 4 ºC for 4hr, equilibrated for 2hrs at 4ºC and packaged into 0.5 ml French straws (20 x 106 spermatozoa/straw). All semen straws were placed into automatic programmable freezer having liquid nitrogen vapors for 10 min. Afterward, shifted to liquid nitrogen for freezing and were stored until post-thaw semen evaluation carried out. The experiment was
repeated for seven times (replicates, n=seven). For post-thaw semen evaluation, four semen straws per treatment group were thawed (30 seconds) in water bath at 37ºC and post-thaw CASA sperm motility parameters and kinematics were checked. Post-thaw motility (PTM%) , Plasma Membrane Integrity (PMI %) , acrosome integrity (AI %) , live Sperm Percentage (LSP % )and sperm abnormalities (SA %) were checked by phase contrast microscope. Similarly AI (%), PMI (%), mitochondrial integrity (MI%) and DNA integrity (%) were checked by fluorescence microscope at Department of Theriogenology, University of Veterinary and Animal Sciences, Lahore.
Pre-freeze CASA sperm motility parameters;progressive %, rapid %and kinematics;average path velocity (VAP um/s), straight line velocity (VSLum/s), linearity (LIN %) were significantly better in Triladyl thanTCEY.Post thaw CASA sperm motility parameters; motile %, progressive %, rapid % and kinematics; VAP (um/s),VSL (um/s), straightness (STR %) and LIN (%) were also significantly better in Triladyl than TCEY. Post thaw semen quality parameters containing PTM (%), PMI (%), LSP (%), DI (%) andMI (%) were significantly better in Triladyl as compared to TCEY.
Availability: Items available for loan: UVAS Library [Call number: 2785-T] (1).
28.
Effect Of Β-Carotene And Tocopherol On Pregnancy Rate In Cidr Synchronized Nili-Ravi Buffaloes
by Muhammad Bilal Munawar (2015-VA-16) | Prof. Dr. Mian Abdul Sattar | Prof. Dr. Nasim Ahmad | Dr. Muhammad Ijaz.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Buffalo is of unique importance in livestock and dairy industry of Pakistan due to its high milk production and shares 65% of total milk production. Reproduction is important to get good production and profit in dairy sector. Reproduction in buffalo is compromised due to its small size ovaries, poor ovarian reserves and less pronounced estrus intensity, resulting as low fertility. Synchronization techniques including CIDR based protocols are well established in cows and getting popularity in buffaloes but with low results, comparatively. Therefore, some modifications are required based on physiology of estrus cycle in buffalo. This was hypothesized that additional injection of Dalmavital in CIDR base protocol will enhance the pregnancy rates and embryonic liability by minimizing the oxidative stress. Therefore, present study is conducted to evaluate the effect Dalmavital on estrus response, Estrus intensity, Pregnancy rates and embryonic losses in CIDR synchronized Nili-Ravi buffalo. For this, buffaloes were scanned ultrasonically for the reproductive tract evaluation. Reproductively sound buffaloes were selected and randomly allocated to one of the two treatment group. 86 buffaloes with normal reproductive tract were assigned in two groups 1; CIDR group (n = 43) and 2; CIDR+D (n = 43). AI was performed twice at 48 and 60 hours after CIDR removal. Estrus response (ER) did not differ significantly (P >0.05) in groups, CIDR and CIDR-D but estrus intensity (EI) was statistically significant (P<0.05) in treatment group. Pregnancy rates were also non-significant (P>0.05) in treatment and control group but improved comparatively in CIDR-D group (63% in CIDR-D group and 56% in CIDR group). Embryonic and fetal losses were also non-significant (P>0.05) between the control and treatment group. Results were also compared in cyclic and non-cyclic, Milking and dry, BCS and parity. Results were non-significant in milking and dry, BCS and Parity. Pregnancy rates were found different significantly (P<0.05) in cyclic and non-cyclic animals, when treatment
Summary
27
is ignored. From the present study it can be concluded that Dalmavital may have good effect on estrus intensity in CIDR synchronized Nili-Ravi buffalo.
Nili-Ravi buffalo is known as black gold of Pakistan. They produce about 2500 liters of milk with 6.5% butter fat. Despite of benefits, this breed is highly influenced with low reproductive activity that include prolonged pubertal period, poor exhibition of estrus, inadequate ovarian activity, long calving interval, high embryonic mortality and low fertility rate with artificial insemination. These factors reduce buffalo’s reproduction which leads to great economic losses. Therefore, there is dire need to address these problems and to orchestrate novel approaches to enhance the reproductive efficiency of buffalo. In cows, during last six to decades, researchers have considerably devised certain strategies to synchronize estrus with the help of prostaglandins, progestagens and estrogens. The advent of these hormones or synchronization protocols helped significantly in improving reproductive management. Moreover, the use of fixed time artificial insemination protocols resulted in acceptable fertility. However, these tools provide inconsistent results to manage reproduction in buffalo. Therefore, unprecedented approaches are required to facilitate and improve the buffalo reproduction. Availability: Items available for loan: UVAS Library [Call number: 2803-T] (1).
