1.
A Quantitative Investigation Of Primordial Follicles In Noncycling 3-4 Years Nili-Ravi Buffalo Heifers
by Izaz Anjum | Rashid Ahmed Chaudry | Imtiaz Hussain Khan | Manzoor | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 1991Dissertation note: Late puberty is the major factor in buffalo heifers leading to low reproductive efficiency resulting in economic losses. Very little information was available about the reproductive potential based on quantitative investigation of primordial follicles and their relation with the ovarian size and weight which directly affects the attainment of puberty.
The present study was designed to determine the number of primordial follicles in non - cycling 3 - 4 years buffalo heifers and results were based on the specific selection of the animals after confirmation of age by dentition, general physical examination, the reproductive status after slaughter. This was achieved by the collection of 12 samples (ovaries) from six animals, two animals belonging to each age group of 3 years, 3 1/2 years and 4 years.
In the heifer Al, the right ovary was more round in shape than left but somewhat softer in consistency than the left ovary. In the heifer A2, the left ovary was somewhat flattened than right ovary which was firmer in consistency than the left ovary and surface was nodular in both ovaries. In the heifer Bi the left ovary was less oval than the right one but was harder in consistency than right ovary. In the heifer B2 the left ovary was more oval in shape than right and somewhat nodular and softer in consistency than right ovary. In the heifer Cl, the left ovary was oval and right elliptical in shape having smooth surface throughout while the right ovary was harder in consistency than left one. In the heifer C2, both ovaries were almost similar in shape which were more oval almost round and right ovary was quite hard and its palpation was almost impossible.
The average weights of ovaries in two non cycling buffalo heifers of Group A (3 years) were 3.30 ± 0.50 gm for the left and 3.90 ± 0.10 gm for the right ovaries (Table 3). The average weights of ovaries in two non cycling buffalo heifers of Group B (3 1/2 years) were 2.70 ± 0.30 gm for the left and 2.90 ± 0.30 gm for the right ovaries (Table 3). The average weights of ovaries in two non cycling buffalo heifers of Group C (4 years) were 2.40 ± 0.40 gm for the left and 2.70 ± 0.70 gm for the right ovaries (Table 3).
The average size of the ovaries of Group A heifers (3 years) was 2.61 ± 0.53 cm x 1.64 ± 0.40 cm x 1.53 ± 0.13 cm in length, width and thickness respectively for the left ovaries and same for the right ovaries was 2.61 ± 0.35 cm x 1.88 ± 0.10 cm x 1.65 ± 0.01 cm (Table 5). The average size of the ovaries of Group B heifers (3 1/2 years) was 2.39 ± 0.13 cm x 1.58 ± 0.10 cm x 1.31 ± 0.13 cm in length, width and thickness respectively for the left ovaries and same for the right ovaries was 2.39 ± 0.05 cm x 1.55 ± 0.21 cm x 1.34 ± 0.09 cm (Table 6). The average size of the ovaries of Group C heifers (4 years) was 2.18 ± 0.08 cm x 1.41 0.16 cm x 1.35 ± 0.0cm in length, width and thickness respectively for the left ovaries and same for the right ovaries was 2.56 ± 0.50 cm x 1.48 ± 0.16 cm x 1.26 + 0.02 cm (Table 7). The mean volume i.e. length x width x thickness for the left and right ovaries of the heifers belonging to Group A was 6.39 ± 0.66 cm3 and 8.05 ± 0.71 cm3 respectively and the same for the heifers belonging to Group B was 5.03 ± 1.08 cm3 and 4.99 ± 0.90 cm3 respectively and for the heifers belonging to Group C was 4.18 ± 0.63 cm3 and 4.86 ± 1.38 cm3 respectively.
The average number of primordial follicles for the left ovaries was 22108 ± 1099 and the same for the right ovaries was 22920 ± 106 in the heifers belonging to Group A (Table 25). The average number of primordial follicles for the left ovaries was 21105 ± 1304 and the same for the right ovaries was 21956 ± 1835 in the heifers belonging to Group B (Table 26). The average number of primordial follicles for the left ovaries was 1620 ± 282 and the same for the right ovaries was 1231 ± 53 in the heifers belonging to Group C (Table 27). In the ovaries of heifers belonging to Group A the mean diameter of the oocyte nuclei was 12.0 ± 1.15u, where as the same in the Group B and Group C were 11.04 ± 1.05u and 11.00 ± 0.02u respectively. The total number of growing follicles in both left and right ovaries of the heifer Al was 123 and the same number of the heifer A2 was 175 with an average of 149 ±26 in both the ovaries of heifers Al and A2 belonging to Group A (Table 28).
The total number of growing follicles in both left and right ovaries of the heifer Bi was 128 and the same of the heifer B2 was 152 with an average of 140 ± 12 in both the ovaries of heifer Bi and B2 belonging to Group B (Table 28). The total number of growing follicles in both left and right ovaries of the heifer Cl was 47 and the same of the heifer C2 was 15 with an average of 31 ± 16 in both the ovaries of heifers C1 and C2 belonging to Group C (Table 28).
The number of primordial follicles was counted per ovary and compared with the relative size and weight of ovary and age of the heifer a poitive corelation was found to be existed between ovarian size/weight and primordial follicle number perovary of heirfers of three age groups and hence the possible factors responsible for culling or selection of such animals were observed for the improvement of buffalo dairy herd.
CONCLUSION and SUGGESTIONS
It is concluded from the results of present study that the ovarian weight and size have a significant effect On the number of primordial follicles since the weight cannot be assessed through rectal palpation and when the average ovarian weight and size were analyzed statistically a +ve correlation of 0.94 was found between these two factors. Hence it is concluded that the normal ovarian weight can he compared with the normal ovarian size while performing the examination of genitalium per rectum and hence by measuring the ovarian size per recturm in heifers its reproductive potential i.e. the number of primordial follicles per ovary can be estimated which would he helpful in selecting or culling virgin buffalo heifers who have not attained puberty. While performing the specific genital examination it is suggested that the following parameters must he considered.
1) The age of heifer for selection must he upto 3 1/2 years.
2) Per rectal examination it must he confirmed that the genitalium has no any segmental aplasia.
3) The ovaries must he oval in shape with somewhat nodular surface, soft, flexible consistency.
4) The ovarian size must he 2.61 + 0.53 x 1.64 ± 0.40 x 1.52 ± 0.12 cm for the left ovaries and 2.61 ± 0.35 x 1.88 ± 0.10 x 1.65 ± 0.01 cm for the right ovaries in length, width and thickness respectively.
5) The ovaries must never have any tumour which can he differentiated from the developing functional structure i. e. follicles and corpus luteum and if the ovarian tumour are confused with functional structures then these animals may he kept under observation for 21 days to find any developmental stage of functional structure.
6) Probably the heifers approaching puberty or in some cycling stage may have 3 - 8 transverse wrinkles upon their external vaginal lips.
It is anticipated that these conclusions and suggestions will serve a useful purpose in selecting or culling those buffalo heifers which have not attained puberty till 3 to 4 years of age and hence will be helpful to make necessary improvements in buffalo dairy herds.
Availability: Items available for loan: UVAS Library [Call number: 0232,T] (1).
2.
Studies On Productive Ability & Fertility Trials Of Standard And Routine Donors Buffalo Bulls Kept At Semen Production
by Ali Raza, Capt. M | Dr . Rashid Ahmad Chaudhry | Dr . Haji Ahmad | Dr . Muhammad Aleem | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 1999Dissertation note: The mature breeding buffalo bull at the Semen Production Unit has greater importance to produce maximum doses and higher fertility rate in the field. The sire should not only be of high genetic make-up but should also be in good sexual health for the production of large number of fertile germ cells based on seminal characteristics.
The present project was designed to investigate the production potential and reproductive performance (fertility trials) of the donor buffalo bulls. On the basis of available data the investigations were carried out in two groups comprising of standard bulls and routine donors. The standard bulls were offered mixed bull ration while routine donors were kept on routine management and available fodder regime.
It is evident from the results that standard buffalo bulls had better and encouraging performance. The mean values of all the seminal parameters among different age groups were comparatively higher .in the bulls fed on additional mixed bull ration than the group of bulls kept on green fodder and routine maintenance ration only.
Although superior seminal values were obtained but statistically the differences within age groups were found to be nonsignificant for both feeding regimes. It may be due to limited duration. However, within various seminal characteristics highly significant differences were observed. After evaluating the semen in the laboratory it is of utmost importance to test this semen in the field conditions because the ultimate determinant of the semen fertility is the conception rate. A total of 3807 doses of semen from ten bulls were distributed in 10 selected A.I. centers. The first insemination resulted in an average fertility rate of 56.52±3.97% whereas the fertility rate of second insemination was 60.37%. The average fertility resulted from all bulls were 56.52±3.97%. Among the bulls the fertility rates were non significantly different. Similarly, mean fertility rates from each A.I. center were not significantly different. It was also observed that there was no significant difference in the fertility rates among various months of semen collection and insemination. The fertility results are better in aged buffaloes when compared to those of heifers (57.99 Vs 54.74%).
Similarly, to compare the effect of three diets, having crude proteins and digestible nutrients from different sources on seminal attributes had indicated better results for implementation at the Semen Production Unit. Seasonality month of the year affected all the seminal attributes. Values of the winter months (November to January) were better than the other three seasons. October was the best month, while the June had worst effect on the semen quality.
Availability: Items available for loan: UVAS Library [Call number: 0696,T] (1).
3.
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).
4.
Effect Of Timing Of Artificial Insemination In Relation To Ovulation On Pregnancy Rate In Sahiwal Cows
by Waqas Ahmad (2007-VA-99) | Prof. Dr. Nasim Ahmed | Prof. Dr. Manzoor Ahmad | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Sahiwal cow is the best milch breed among all Bos indicus throughout the Subcontinent. It is famous for heat resistance, tick tolerance, high milk production. It is also exported to different countries for its peculiar properties. These cows have low fertility when inseminated with frozen semen. Very little fragmented experimental data is available on reproduction of Sahiwal breed, which might be major reason of bad performance from reproduction aspect. The same rule of insemination as we know in Holstein (Bos taurus) when applied on our local animals did not yield similar results. The designed experiment was to determine appropriate time of insemination to improve reproductive efficiency in Sahiwal cows.
This experiment was conducted at Livestock Experimental Station Jahangirabad, District Khanewal Punjab. Eighty five (n=85) adult, multiparous, lactating Sahiwal cows having BCS ≥ 2.5 and weight 325–450 kg, more than 60 days post-partum with normal reproductive tract were selected for this study. Estrous detection was carried out twice daily with teaser bull for 30-45 minutes. Standing heat was considered when cow did not move away for 4-6 seconds with teaser bull being mounted on her. Cows were assigned randomly into four groups (0 h, 12 h, 24 h and 36 h) with respect to standing heat. Frozen semen from bull whose fertility is known to us having post thaw motility of 40% at least was used for insemination.
Ultrasound was used as tool for precise assessment of reproductive status of experimental animals. At the start of each replicate, both ovaries and uterus of all the adult Sahiwal cows were scanned with B-Mode Ultrasound console for presence or absence of fetus and CL or any structural abnormality, with help of 7.5MHz Trans-rectal probe. Pregnancy was diagnosed 35 day post AI. Results were analyzed by using statistical software (SPSS). Pregnancy per AI was compared amongst
CHAPTER 6
SUMMARY
Summary
37
different insemination groups by using binary logistic regression test. Pregnancy was assessed retrospectively by plotting scatter graph using ovulation as our reference point.
The timing of ovulation was 20.64 hours from onset of standing heat. Mean size of ovulatory follicle at 0, 12, and 24 hour after standing heat was 13.52 mm, 14.52 mm and 15.39 mm respectively. The ovulation rate was 97%. Highest pregnancy per AI 57% (13/23) was observed in 0 h group, followed by 36% (8/22) in 12 h, followed by 25% (5/20) on 24 h group. Lowest pregnancy per AI 10% (2/20) was seen in group inseminated 36 hour after onset of standing heat (36 h). While retrospectively highest pregnancy per AI 67% (10/15) was observed in -18 h group, followed by 29% (6/21) in -6 h group and 32% (7/22) in +6 h group, while no pregnancy were observed 0% (0/12) in +18 h group. The overall pregnancy per AI was 33% (28/85) in Sahiwal cows. Availability: Items available for loan: UVAS Library [Call number: 2468-T] (1).
5.
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).
