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1. 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: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1900,T] (1).

2. 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: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2005,T] (1).

3. 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: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2018,T] (1).

4. 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: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2085,T] (1).

5. 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: book Book; 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 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).

6. Epidemiological Studies And Chemotherapy Of Postparturient Haemoglobinuria In Buffaloes

by Muhammad Ijaz Ul Haq (2008-VA-83) | Dr. Jawaria Ali Khan | Dr. Muhammad Avais | Prof. Dr. Mian Abdul Sattar.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: The district Gujranwala lies at northern part of the Punjab in Pakistan. It is situated between two main cities, in North is Gujrat and in South, there are Sheikhupura and Lahore. The majority of the population lives in the rural areas and mainly earns their livelihood through agriculture. The source of irrigation is perennial and non-perennial canals supplemented by tube wells. This area is heavily populated with buffaloes rather than cattle. Buffalo is playing a foremost role in the national economy by producing milk, meat and draught power. Out of total milk produced in the country, buffalo contributes about 68 %. Due to high fat contents of buffalo milk, it is the ideal species in Pakistan (Bilal et al. 2006). Buffaloes suffer from many fatal diseases throughout the year causing irretrievable economic losses in the GDP. Among these fatal diseases, Postparturient haemoglobinuria is a frightening problem for the economy of Pakistan and as well as for the whole world. Postparturient haemoglobinuria is a periodic condition seen worldwide that most frequently affects high-yielding dairy animals at the onset of lactation. It is characterized by development of per acute intravascular hemolysis and anemia with potentially lethal results. Beef and non-lactating cattle are hardly ever affected. The exact cause is unidentified, but phosphorus reduction or hypophosphatemia is known to be a major predisposing factor. Decrease in the Phosphorus level of RBCs is known to raise osmotic tenderness of the RBCs, probably predisposing to intravascular hemolysis. A parallel condition reported in New Zealand was linked with copper depletion, potentially building RBCs were more prone to oxidative trauma. Other potential causes were hemolytic or oxidative plant toxins (Michael A. Moses and Aiello 2014). In buffalo rearing countries mainly in India, Pakistan and Egypt, Postparturient haemoglobinuria was a major problem as described by (Pirzada and Hussain 1998). Acute hypo phosphataemia was linked with high producing buffaloes and characterized by intravascular hemolysis (Kurundkar et al. 1981). Postparturient haemoglobinuria was a disease of high yielding buffaloes in district Attock, Rawalpindi, Jhelum and Faisalabad where soil was deficient in mineral contents. The disease was distinguished by icteric and inflexible constipation in pregnant animal (Hussain et al. 1991). Postparturient haemoglobinuria was mainly disease of buffaloes and found in almost all the districts of Pakistan (Pirzada and Hussain 1998). In buffaloes, Postparturient haemoglobinuria occurred mostly after 3rd week of parturition during 3rd to 5th lactation having a period of 5 to 6 days of illness (Nagpal et al. 1968). The occurrence of hypophosphatemia was highest in buffaloes in progressive pregnancy or in 3rd to 6th lactation (Gautam et al. 1972). Multiparous females were mostly affected by Hypo phosphataemia and a range of etiological factors were reported to be related with the disease in unusual parts of the world (Singari et al. 1991). Dietary phosphorus shortage and rations containing cruciferous plants were supposed causes of rigorous hypo phosphataemia and had been associated with hemolytic anemia in cows and buffaloes (Heuer and Bode 1998). Mucous membranes turned pale or might become icteric with the development of anemia and breathing became rapid and shallow with tachycardia (Spears 2003). During third to sixth lactation of adult dairy buffaloes, this disease was more frequently seen, showing weakness and recumbency (Macwilliams et al. 1982). The occurrence of the disease in the total cattle population was very stumpy with a case fatality rate ranging from 10% to 50% (Madsen and Nielsen 1939). Sporadically, farms with a predominantly elevated incidence might be encountered but typically only one or two cases were clinically evident at one time (Freudenberg 1955). In Australia, about 70% cases were reported within 30 days of calving (Mulijns and Ramsay 1959). Bulls, ante partum cows, heifers less than two years old and beef cows were rarely affected. A disease called as "red water" in Ontario, was allied with numerous predisposing factors which integrated recent parturition, intense milk production, dietetic phosphorus absence, utilization of turnips, rape, kale, green alfalfa and sugar beet pulp (Macwilliams et al. 1982). Postparturient haemoglobinuria was one of the foremost and economically important diseases of dairy animals in Pakistan, India and elsewhere in the world (Macwilliams et al. 1982; Chugh et al. 1996; Pirzada and Hussain 1998). Diagnosis was done on the basis of clinical signs, urinalysis and hematological reports of the examined animals (Madsen and Nielsen 1939). In a survey conducted during 1996 in Punjab (Pakistan), mortality rate due to Postparturient haemoglobinuria in buffaloes and cattle was 5% which translated into estimated annual losses of Rs. 490.2 and 153.1 million, respectively (Farooq et al. 2007). Availability: Items available for loan: UVAS Library [Call number: 2210-T] (1).

7. 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: book Book; 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).

8. 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: book Book; 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).

9. 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: book Book; 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).
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10. 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: book Book; 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).

11. 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: book Book; 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).

12. 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: book Book; 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).

13. 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: book Book; 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).

14. 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: book Book; 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).

15. 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: book Book; 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).

16. 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: book Book; 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).

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