EFFECT OF SEASON AND MANAGEMENT ON SEMEN QUALITY OF BREEDING BULLS- A REVIEW

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1 Agric. Rev., 30 (2) : 79-93, 2009 AGRICULTURAL RESEARCH COMMUNICATION CENTRE / indianjournals.com EFFECT OF SEASON AND MANAGEMENT ON SEMEN QUALITY OF BREEDING BULLS- A REVIEW M. Bhakat, T.K. Mohanty, A.K. Gupta and V.S. Raina National Dairy Research Institute, Karnal, Haryana , India ABSTRACT Artificial insemination is nowadays a widely used technique for the enhancement of genetic potential of livestock species. But still the maximum benefit of this tool cannot be extracted as there are certain inherent problems of semen quality like poor semen quality and poor freezability with crossbred and. Correspondingly, there is a genetic and economic loss owing to disposal of these high genetic merit. Semen quality is affected by several factor of which season is one of the major factors. It exerts its effect on reproductive performance through macro and micro climatic factors like temperature, humidity, rainfall, photo-period. On the other side vaccination of exotic and crossbred animals for prevention of contagious diseases, as they are more prone to them in turn has detrimental effect on the semen quality. Therefore suitable management strategies are being searched for and incorporated, like laboratory procedures i.e. the use of filtration technique so as to maximize quality semen production without discarding too many poor quality ejaculates and re-scheduling of vaccination protocol to unfavorable season in terms of semen production. Key words: Season effect, Semen quality, Bulls. The technique of artificial insemination using the cryopreserved semen is presently widely used all over the world to increase the genetic potential of livestock species. Several attempts are being made to increase the AI coverage area so as to obtain maximum out of this technology. But there are certain hindrances in this path of attaining breeding goals. These are the inherent and functional constraints of rearing, which include an overall weak and erratic libido. The characteristically possess this trait. Besides this certain anatomical and physical limitations eg. testicular size is relatively smaller, daily sperm production rate is lower and also the epididymal sperm reserve is less are also there. (Suryaprakasam and Rao 1993, Sudheer and Xavier 2000, Singh 2003). On the contrary, cattle do not have these kind of natural constraints as do. But similar inherent problems pertaining to semen quality are also observed with crossbred (HF crosses). Apart from the inherent traits, season is one major factors which influences the reproductive performance of these animals and it exerts its effect through macro and micro climatic factors like temperature, humidity, rainfall, photo-period (Mandal et al. 2000). Another stress factor is the use of vaccines to prevent them from various contagious diseases and this has shown to have an adverse effect on quality of semen leading to a number of sperm abnormalities (Kammar and Gangadhar 1998, Murugavel et al. 2000, Mathur et al. 2003). Adverse climatic condition, post vaccination stress and treatment for any type of pyrexia and other conditions in due time show severe results. It may result in disposal of those and crossbred which show poor semen quality (those producing consistently below 60% motility of semen) (22% and 45.42%); poor freezability (consistently giving below 35% post thaw motility) (4.1% and 10.87%) and poor libido (11.04% and 36.46%) (Suryaprakasam and Rao

2 80 AGRICUTURAL REVIEWS 1993, Sudheer and Xavier 2000). This becomes a cause for huge wastage and consequently results in huge economic losses, as these are discarded at around 2-1/2 and 3 years of age. Apart from this, there is also a reduction in the genetic gain due to disposal of selected for breeding purpose. At certain time even the possessing good quality semen with good freezability and as well as good libido also tend to produce low grade ejaculates under such stressful conditions. Therefore, the trend now is to design procedures or refine methodologies so as to maximize the production of good quality semen without discarding too many poor quality ejaculates. As of now numerous techniques have been worked out for reducing the number of dead and abnormal cells in the semen as these dead and abnormal sperm have an adverse affect on companion normal cells fertilization potential by the release of certain toxic substances (Shanon 1972). These techniques include percol density gradient (Parrish et al. 1995), swim up procedure (Hallap et al. 2004), glass wool filtration (Vyas et al. 1992, Ulgen et al. 2004), sephadex gel filtration (Vyas et al. 1992, Ahmad et al. 2003) and sephadex ion- exchanger filtration (Anzer and Graham 1993, Ahmad et al. 2003). For sperm recovery several centrifugation techniques (Knop et al. 2005) have been used, which showed considerable improvement in poor quality ejaculates. There are certain reports regarding recovery of sperm after filtration through sephadex showing improved post thaw quality in buffalo (Chauhan et al. 1993, Goyal et al. 1996). Disposal of breeding The major reasons for disposal of breeding are the poor semen quality in crossbred and murrah buffaloes (Mathew et al. 1984, Suryaprakasam and Rao 1993, Sudheer and Xavier 2000) and the poor semen freezability in case of Exotic and (Kotayya and Rao 1981 in Jersey and Ongole, Mathew et al in Brown swiss, Suryaprakasam and Rao 1993 in Jersey, Holstein and Jersey Zebu, Rao et al in Jersey, Jersey x Ongole and Ongole, Sudheer and Xavier 2000 in ). Sethi et al. (1989) and Khate (2005) reported that disposal of Karan Fries and was mainly because of poor quality of semen production. Kodagali et al. (1980) reported culling of 23.81% Surti due to poor semen quality. Chenoweth et al. (2003) also reported from their studies on 98 that one of the major problems of culling encountered in was poor semen quality (4%), where, poor semen quality means the semen samples having less than 60 per cent individual motility and poor semen freezability means the semen samples showing post thaw motility less than 35 per cent. According to study of Khate (2005), regarding production of breeding at NDRI herd (1991 to 2000), it has been found that out of total male calves born % KF and % calves were reserved for breeding purpose on the basis of dam s milk yield or pedigree performance. Out of total male calves reserved only % KF and % buffalo produced semen, whereas others were disposed of due to various reasons. The semen of only % KF and % was found to be freezable. From economic point of view, this led to huge economic loss. At NDRI herd breeding are culled due to various reasons among them poor semen quality, which contribute % in case of KF and 4.76 % in case of buffaloes are the major reasons of culling. Disposal of breeding due to poor semen quality and poor semen freezability have also been reported by Suryaprakasam and Rao (1993) in Andhra Pradesh in case of buffalo (22 % and 4.1 %) and Sudheer and Xavier (2000) in Kerala in (45.42 % and %). Mathur et al. (2002) reported from their observation of 302 Frieswal that percent initially had very poor semen quality i.e. not upto the minimum standard required for semen freezing and only percent yielded freezable quality semen per cent produced satisfactory initial semen quality but had poor freezability due to poor post thaw revival. Tyagi et al. (2006) conducted studies to observe impact of various levels of HF inheritance and on number of indigenous breed components (Sahiwal, Red

3 Vol. 30, No. 2, Significant Seasonal Effect Ejaculate volume (ml) Mass activity (0 to 5 scale) Sperm concentration ( 106/ml) Species Variations Authors Exotic and Bos brachyceros Zebu-taurus Zebu-taurus Egyptian Bos brachyceros Exotic and Zebu-taurus Table 1: Significant seasonal influence on semen quality Dry (Nov- Apr): 3.42 Rainy (May- Oct):5.8 Dry (Dec- Mar): 1.8 Rainy (Apr- Aug): 2.3 Late rainy (Sep- Nov): 2.0 hot-dry(may- June): 3.70 hot-humid (July- Aug): 4.32 Autumn (Oct- Nov): 4.16 Winter (Dec- Jan): 5.04 Summer (May- July): Autumn (Aug- Oct): Winter (Nov- Jan): Spring (Feb- Apr): hot-dry: 3.24 hot-humid: 3.22 Autumn: 3.32 Winter: 3.54 Summer (Apr- June):1.91 Rainy (July- Oct): 1.90 Winter (Nov- Mar): 2.27 Summer: Autumn: Winter: Spring: Summer: Autumn: Winter: Spring: Dry: 216 Rainy: 164 Late rainy: 198 Dry : 970 Rainy : 1742 hot-dry: hot-humid: Autumn: Winter: Summer: Rainy: Winter: Rekwot et al. Igboeli et al. Goswami et al. (1991) Manik and Mudgal (1984) Goswami et al. (1991) Mandal et al. (2000) Oloufa et al. (1959) Manik and Mudgal (1984) Igboeli et al. Rekwot et al. Goswami et al. (1991) Mandal et al. (2000) Cont...

4 82 AGRICUTURAL REVIEWS Live % Hariana buffalo bull Spring (Feb- Apr): Summer (May- July): 67.2 Autumn (Aug- Oct): 68.9 Winter (Nov- Jan): 83.7 Summer (Mar- June): 88 Monsoon (July- Oct): 93 Winter (Nov- Feb): 89 Summer: Autumn: Winter: Spring: Summer Pre-freezing 88 (Mar-May) Post-freezing 66 Rainy (Jun- Pre-freezing 89 Aug) Post-freezing 67 Autumn Pre-freezing 90 (Sep-Nov) Post-freezing 68 Tomar et al. (1966) Dixit et al. (1984) Manik and Mudgal (1984) Bhosrekar et al. (1991) Winter Pre-freezing 88 (Dec-Feb) Post-freezing 67 Motility (%) Zebu-taurus Bos brachyceros Zebu-taurus Surti buffalo hot-dry: hot-humid: Autumn: Winter: Summer: Rainy: Winter: Summer (May- Aug): post thaw motility after 15 min after 30 days Winter (Dec- Mar): post thaw motility after 15 min after 30 days Dry: 36.2 Rainy: 37.7 Late rainy: 27.5 hot-dry: hot-humid: Autumn: Winter: Rainy (Jun- Sep) Winter (Oct- Jan) Initial Motility Post thaw Motility Initial Motility Post thaw Motility Post thaw Motility (Age-1) (Age-2) (Age-1) (Age-2) (Age-1) (Age-2) (Age-1) (Age-2) (Age-1) (Age-2) Goswami et al. (1991) Mandal et al. (2000) Tuli and Singh (1983) Igboeli et al. Goswami et al. (1991) Bhosrekar et al. (1992b) Cont...

5 Vol. 30, No. 2, Summer Pre-freezing 95 (Mar-May) Post-freezing 92 Functional integrity of sperm membrane Rainy (Jun- Pre-freezing 94 Aug) Post-freezing 90 Autumn Pre-freezing 95 (Sep-Nov) Post-freezing 93 Winter Pre-freezing 94 (Dec-Feb) Post-freezing 92 Bhosrekar et al. (1991) Total abnormality %, Bos brachyceros Exotic and Summer: Rainy: Winter: Summer: Autumn: Winter: Spring: Dry: 15.1 Rainy: 9.7 Late rainy: 8.9 Dry : Rainy : 6.45 Summer Pre-freezing 5.7 (Mar-May) Post-freezing 8.8 Mandal et al. (2000) Manik and Mudgal (1984) Igboeli et al. Rekwot et al. Rainy (Jun- Pre-freezing 5.4 Aug) Post-freezing 8.1 Autumn Pre-freezing 5.8 (Sep-Nov) Post-freezing 7.7 Bhosrekar et al. (1991) Winter Pre-freezing 7.3 (Dec-Feb) Post-freezing 9.5 Damaged acrosome% ph Summer: Rainy: Winter: 7.32 Summer: 6.70 Rainy: 4.78 Winter: 1.81 Spring: 6.8 Summer: 6.8 Autumn: 7.0 Winter: 6.9 Mandal et al. (2000) Mandal et al. (2000) Tomar et al. (1966)

6 84 AGRICUTURAL REVIEWS Sindhi, Tharparkar and Haryana) on reproductive wastage rate of the. They concluded that there was an overall 8.36% culling of for not donating semen and only 41.11% (out of those donated semen) produced freezable quality semen. Effect of season The effect of season is both direct and indirect. It affects the animal directly through macro and micro climatic factors, which are the temperature, humidity, rainfall and photo-period. Indirectly it acts by affecting the vegetation, forage quality and soil-plant-animal interaction. Surrounding of a species, including human being, has great influence on its desire for sexual activity and the variations regarding this desire may occur with reference to time, place and the subject concerned. India has different agroechological zones having wide variety of climatic conditions and this can naturally lead to variations in findings of various authors. In general, of all seasons, summer exerts comparatively more adverse effect on the overall quality of semen. It effects the normal reproduction process multidimensionally, like by reducing feed intake, by inhibiting the release or response to important reproduction hormones that is, GnRH, FSH and LH. LH is the important hormone, responsible for spermatogenesis. It is inhibited by increasing level of plasma corticosteroids (Clarke and Tilbrook 1992) due to heat stress. During summer, thyroxin secretion declines leading to reduced intake of feed by the animal, subsequent metabolism (Madan 1985) and is responsible for reducing production of sperm (Zafar et al. 1988). Due to extreme heat stress get physically exhausted and their reduced eagerness might result in higher reaction time and subsequently total time for successful ejaculation also increase, thus having an ultimate effect on production of sperms (Mandal et al. 2000). Series of researches conducted regarding effect of season on semen quality parameters by various workers and few of the m with respect to significant seasonal effect and non significant seasonal effect on semen quality parameters are presented in Table 1 and Table 2, respectively. Besides these series of research, Ravimurugan et al. (2003) reported significant (P<0.01) seasonal influence on ejaculate volume, motility and sperm concentration southwest monsoon proved to be best season in semen volume, cold weather in motility and sperm concentration in. Bhat et al. (2004) studied on 4446 ejaculates over a period of 7 years to know the seasonal influence on various seminal attributes, in. The influence of season on semen volume and mass activity was significantly higher (P<0.05) during monsoon. Sperm concentration was highest in summer. The effect of season on progressive motility was non-significant. The study concludes that monsoon season is most favourable for production of quality semen follows by summer and winter. Koonjaenak et al. (2007) studied on five mature, healthy swamp buffalo AI in Thailand from July 2004 to the end of June 2005, regarding effect of season on semen quality parameters of swamp buffalo. He observed ejaculate volume, ph, sperm concentration, total sperm number and initial sperm motility did not differ between seasons, whereas PMI and the relative proportion of morphologically normal spermatozoa were highest in summer and lowest in winter (P<0.05). Season influenced sperm morphology (P< ). Among morphological abnormalities, only proportions of tail defects were affected by season, being highest in the rainy season and lowest in summer (P<0.01). Helbig et al. (2007) reported that semen collection at seasonal intervals failed to show any changes in sperm cell concentration, volume, density and gross motility in American bison. Mathur et al. (2002) observed that there was no significant seasonal difference in mass motility, progressive motility and sperm concentration except that volume of semen, which was lowest during winter (3.84 vs. 4.17, 4.18 ml), compared to summer and rainy seasons in Frieswal. Singh and Raina (2000) investigated seasonal influence on six crossbred (464 observations) over a period of one year and found that season had significant (P<0.01) effect on ejaculate volume, mass activity, abnormal sperm percent and sperm concentration.

7 Vol. 30, No. 2, Table 2: Non significant seasonal influences on semen quality Non-significant Seasonal Effect Ejaculate volume (ml) Motility (%) Species Variations Authors Egyptian Hariana Egyptian Exotic and Nili- ravi Summer: 3.94 Autumn: 3.26 Winter: 3.29 Spring: 3.48 Spring: 4.4 Summer: 5.0 Autumn: 4.5 Winter: 4.2 Spring: 4.8 Summer: 5.4 Autumn: 5.0 Winter: 5.0 Summer: Autumn: Winter: Spring: Summer: Autumn: Winter: Spring: Dry : Rainy : hot-dry (Apr- June): 63 hot-humid (Jul- Sep): 64 warm (Oct- Nov): 64 cold (Dec- Mar): 64 Summer Pre-freezing 86 (Mar-May) Post-freezing 63 Rainy (Jun- Pre-freezing 86 Aug) Post-freezing 64 Autumn Pre-freezing 86 (Sep-Nov) Post-freezing 63 Winter Pre-freezing 85 (Dec-Feb) Post-freezing 63 Summer: Rainy: Winter: Oloufa et al. (1959) Tomar et al. (1966) Tomar et al. (1966) Manik and Mudgal (1984) Oloufa et al. (1959) Rekwot et al. Zafar et al. (1988) Bhosrekar et al. (1991) Mandal et al. (2000) Cont...

8 86 AGRICUTURAL REVIEWS Uve% Summer: Egyptian Autumn: Oloufa et a/. Winter: (1959) Spring: Hariana Spring: 86.5 Summer: 85.4 Tomar et aj. Autumn: 85.0 (1966) Winter: 83.5 Spring: 82.4 Summer: 67.2 Tomar eta/. Autumn: 68.9 (1966) Winter: 83.7 Hot dry: 2.40 Mass Activity Nili-ravi Hot humid: 2.43 Zafar etaj., (0 to 5 scale) Warm: 2.40 (1988) Sperm concentration (x10 6 /ml) Abnomal sperm% ph Cold: 2.31 hot-dry: 1063 NiH- ravi hot-humid: 949 Zafar et aj. warm: 1006 (1988) cold: 933 Rainy (Jun-Sep) Age Age Surti buffalo Age Bhosrekar et Winter (Oct-Jan) Age aj. (1992b) Summer (Feb-May) Age Age Summer: 8.73 Egyptian Autumn: Oloufa etaj. Winter: (1959) Spring: 9.87 Summer: 6.82 Egyptian Autumn: 6.85 Oloufa etaj. Winter: 6.92 (1959) Hariana Spring: 6.93 Spring: 6.5 Summer: 6.5 Tomar et aj. Autumn: 6.8 (1966) Winter:6.7 Summer: 6.97 Mandai et a/.. Rainy: 6.85 (2000) Winter: 6.87

9 The procedure of semen quality assessment, instruments used, expertise of different evaluator, replications of the experiment and seasonal classification might have resulted in little consistency in results. Effect of vaccination In India, as a regular prophylactic measure, breeding are vaccinated against various contagious diseases, but still more emphasis need to be laid on vaccination of exotic as they are more prone to FMD, HS, BQ and other diseases (Mathur et al. 2003). Semen quality is affected by vaccination (Gahlot and Kohli 1981, Venkatareddy et al. 1991, Murugavel et al. 1997) as it is one of the major stress factors which cause rise in body temperature because of the febrile reaction occuring during post-vaccination period. Also there is derangement in spermatogenesis (Venkatareddy et al. 1991) due to elevated temp of testes and besides this abnormal acrosome development (Bane and Nicander 1966) occurs temporarily due to induction of testicular degeneration. This preventive vaccination might be followed by epididymal dysfunction which possibly is the reason for increased number of spermatozoal abnormalities in the initial stage, resulting in reduced motility (Rao and Venkataswami 1971, Rao 1976). These derogatory effect of vaccination resembles those of therapeutic agents on germinal cells resulting in increased number of dead spermatozoa, which leucocytes absorbs through the process of phagocytosis (Mann and Mann 1981). There is subsequent decline in epididymal sperm reserves (Rao et al. 1980), thus concentration decreases as the resorption of abnormal sperm increase. Very less information is there in concern with the semen quality during post vaccination period. As compared to bacterial vaccine, viral vaccine has more deleterious effects, as both the metabolic activity and cold shock resistance of sperm are reduced to a considerable level (Venkataswami et al. 1972). Semen quality (Saxena and Tripathi 1976) of exotic and crossbred (Tripathi and Saxena 1976, Saxena and Tripathi 1977, Gahlot et al. 1990, Venkatareddy et al. 1991, Mathur et al. 2003) is adversely affected by vaccinations Vol. 30, No. 2, against FMD, HS and BQ. Findings of various workers on these aspects have been summarized in the Table 3. Aversion of pyrexia, so as to maintain normal body temperature can be a preventive measure against the adverse effects of vaccination (Gahlot et al. 1990). The pyretic response to these vaccinations varies from bull to bull, breed to breed and even species to species. To certain extent this adverse effects can also be reduced by immuno-modulators. Therefore, it is advised that vaccination schedule may be followed when the season is unfavorable as during that time the semen production is usually of poor quality especially for purpose of freezing. Effect of collector Semen collection is the business for various Artificial Insemination organizations, hence the management of entire process of collection is quite critical and for this the vital role is played by the semen collector. The amount of motile sperm collected might be influenced by the bull handler,s skill and behavior (El-Wishy 1978,) Since both the motility (Foster et al. 1970) and the number of sperm obtained (Almquist 1973) are enhanced only if the sexual stimulation is appropriate. As perceived from the reports of Mathevon et al. (1998) that in both young (P < 0.001) and mature (P < 0.01) the volume of ejaculate is significantly affected by the variation in collection team. In both the cases, the effect of collection team was significant (P < 0.05) for total number of cells and motile cells per ejaculate. The collection team did not have a significant (P > 0.1) effect on concentration and motility of the sperm. Collection team by treatment interaction were examined by Yates et al (2003), Team 2 was more efficient than Team 1 in collecting the at night rather than during the day with a significantly shorter time to first mount at night. Thus, one can conclude that compare to night, during day time were more of interruptions for the semen collection team. For instance, the commonest of all disruptions were the phone calls for collection team members and co-worker questions from people not involved in the semen

10 88 AGRICUTURAL REVIEWS Table 3: Effect of vaccination on semen quality Breed Findings Authors Decline in motility, concentration and percentage of live spermatozoa during post-vaccination period before returning to normal levels. Following Rinderpest vaccination marked decline in in-vitro semen preservability. Radhakrishnan et al. (1975) During 0-45 days post-vaccination of FMD the abnormalities of sperm increased and semen preservability declined. Tripathi and Saxena (1976) Jersey Jersey Exotic, and zebu Frieswal Buffalo Sunandini Karan Fries (KF); (MU) and Sahiwal (SW) Motility, livability, preservability of sperm declined and abnormalities increased upto 45 days post-vaccination of FMD. Mild deleterious effects of RP vaccination were observed as compared to reports by Radhakrishnan and coworkers in Pyrexia was not noticed post vaccination. FMD vaccination effect in Jersey and Jersey Ongole was moderate, but in ongole it was less. 3 wks following vaccination the initial motility and live sperm percentage declined and total spermatozoal abnormalities increased significantly. Semen quality parameters showed a declining trend: mass activity, from 3.1 to 2.5 and mean per cent progressive motility 55.4 to 46.5 at pre FMD and post-hs and BQ vaccination, respectively. On HS and BQ vaccination had more detrimental effects. The adverse effects on the semen quality (concentration, motility, live sperm count, intact acrosome and host decrease) as exerted by the FMD vaccination existed upto 1 month after vaccination. In Kerala Livestock Development Board also semi-annual vaccination against Foot & mouth disease followed with two weeks sexual rest was done to cope with post vaccination stress. Application of Raksha Triovac vaccine had adverse effect on sperm motility and mass activity of MU and KF bull semen. Vaccination had no significant (P>0.05) effect on volume and sperm concentration of semen in KF, MU and SW bull semen. Saxena and Tripathi (1977) Gahlot and Kohli (1981), Gahlot et al. (1990) Venkatareddy et al. (1991) Murugavel et al. (1997) Mathur et al. (2003) Singh et al. (2003) Krishnan et al. (2003) Pankaj (2006)

11 Vol. 30, No. 2, Breed Findings Authors Holstein Improved percentage of motile sperm after freezing and Graham and Graham thawing after sephadex filtration (G-15). lower fertility (1990) showed improvement of 60 to 90-d nonreturn rates. Improvement in sperm motility and quality following the use of glass-wool of sephadex G-10 column filtration over the whole semen and it could then be preserved at 5 0 C and satisfactory sperm motility observed upto 96 hrs only. Significant improvement in sperm motility, live sperm per cent, abnormal per cent and abnormality could be achieved by filtration through glasswool and sephadex (G-25, G-50), particularly G-25. Use of G-75 sephadex (G-25, G-50, G-100, G-200) columns showed significant improvement of sperm motility, live sperm per cent, per cent intact acrosome as compared to unfiltrated control. Holstein Different kinds of immotile spermatozoa from an extended ejaculate can be removed successfully by use of sephadex ion-exchange column. Better post thaw viability was observed. Buffalo Holstein Nili-Ravi Initial semen quality improved. More effective yield of freeze processed semen was obtained with simple washing procedure and then suspension in a suitable extender as compared to that obtained following sephadex filtration and freezing. A higher number of post-thaw normal acrosome and instances of intact plasma membrane as well as higher swelling rates and better in vitro penetration ability of ZFHO was then with sperm Filtered by sephadex ion-exchange column. In harvesting maximum number of motile spermatozoa from frozen-thawed semen (95 vs 33%; P<0.001), the Ionexchange filtration proved better than swim-up procedure. Buffalo Semen filtration through sephadex column (G ) showed significantly (P<0.05) better maintenance of sperm motility, live sperm count, intact acrosomes and sperm morphology. Buffalo and Holstein Table 4: Effect of filtration on semen quality The semen frozen after filtration (sephadex G-200 and glass wool) showed significantly (P<0.01) higher progressive sperm motility, live spermatozoa and intact acrosomes at various stages of freezing were in. FS+IE columns filtration had high percentages of motility, normal acrosome and intact plasma membranes as compared to FS and control and showed low percentages of sperm abnormalities at all stages of cryopreservation. The proportion of motile, live sperm having improved osmotic resistance and penetrating ability increased with sephadex (G-15) filtration of post thaw semen. Vyas et al. (1991) Vyas et al. (1992) Chauhan et al. (1993) Anzar and Graham (1993) Anzar and Graham (1995) Anzar and Graham (1996) Goyal et al. (1996) Anzar et al. (1997) Mustafa et al. (1998) Kumar et al. (1999) Panghal and Tuli (1999) Ahmad et al. (2003) Kumar et al. (2003) Continu...

12 90 AGRICUTURAL REVIEWS Gir and Jafarabadi Dairy The was increase in per cent progressive sperm motility live sperm and decrease in per cent abnormal spermatozoa and damaged acrosome in sephadex G-100 filtered semen ascompared to non-filtered frozen thawed semen ascompared to non-filtered frozen thawed semen Sephadex G-100 and G-200 columns proved to be more efficient than others (G-25 to G-75) with regards to overall improvement of quality of post-thaw semen. Significantly reduced the concentration of recovered spermatozoa (P<0.01), but improved semen quality, reduced the number of spermatozoa with various forms of morphological defects after sephadex (G-15) filtration. Filtration also affected percentages of sperm motility after equilibration and after freezing/thawing. Maurya and Tuli (2003) Rana et al. (2004) Januskauskas et al. (2005) collection process, on the other hand night time had an upper edge as only people that were present at the arena were semen collection team members and because of the late hour outsiders disturbances were also nonexistent. Kerruish (1955) also reported that bull s reaction can also be affected by several extraneous factors like attendants and nearby structures. Motivation of collector and handler for their work is necessary to optimize semen production and greater incentives need to be given to workers to improve their techniques (Prosser 1968, Hafs 1972, Amann and Almquist 1976, Larson et al. 1982) Effect of filtration In the semen of poor quality, numerous dead and abnormal sperms are present which release ROS and certain other toxic substances. Toxic effects of ROS on sperm was earlier reported in 1943 by MC Loed and it is thought to have an adverse effect on normal companion cells fertilization potential (Shanon 1972). ROS acts through lipid peroxidation of carbon chain of unsaturated fatty acid, which are the important constituent of sperm plasma membrane and thereby results in formation of highly cytotoxic lipid hydroperoxides. These lipid hydroperoxides decompose to form end product malondialdehyde, which is highly toxic and is responsible for DNA and protein damage finally leading to cell death. ROS Production though can not be prevented to full extent but can still be minimized by taking up the following measures; these include Percol density gradient (Parrish et al. 1995), Swim up procedure (Hallap et al. 2004), glass wool filtration (Vyas et al. 1992, Ulgen et al. 2004), Sephadex gel filtration (Vyas et al. 1992, Ahmad et al. 2003) and sephadex ion- exchanger filtration (Anzar and Graham 1993, Ahmad et al. 2003). For sperm recovery various centrifugation techniques (Knop et al. 2005) have also been used. Table 4. summarizes the reports on effect of filtration on semen quality. According to the available literature, there is a clear indication that after filtration of sperm through Sephadex, the post thaw quality of spermatozoa recovered is improved than control in (Chauhan et al. 1993, Goyal et al. 1996). Though the mechanism behind the trapping of spermatozoa in a sephadex column is still not clear, but it has been hypothesized that the Sephadex particle can act like a barrier. It either allows agglomeration of immotile or dead spermatozoa (Graham et al. 1976) or the protein present on the plasma membrane of capacitated sperms binds to the Sephadex particles and finally leading to entrapment of these sperms (Samper, 1990). In case of Sephadex-ion-exchange filtration, it is hypothesized that positively charged components show appearance on surface of dead sperm cell and they can further interact with negatively charged carboxy methyl

13 cellulose (CM- 52), resulting in trapping dead spermatozoa. From these comparative findings, we can conclude that sephadex with ion exchange filtration can facilitate the use of even low grade ejaculates of high genetic. So it is the need of the hour to integrate these techniques in freezing protocol for augmenting the germplasm production. For maximizing quality germplasm production the following critical control points need to be focused: Vol. 30, No. 2, Suitable managemental intervention for amelioration of seasonal stress specially heat stress must be introduced. Vaccination protocol may be rescheduled shifting it preferably to unfavourable season in terms of semen production. Semen collector must be properly trained and highly motivated In case of poor quality ejaculates Sephadex with ion exchange filtration can augment the quality germplasm production. REFERENCES Ahmad, Z. et al (2003). Theriogenolo, 59: Almquist, J. O. (1973). J. Anim. Sci. 36: Amann, R. P. and Almquist J. O. (1976). In: Proc. 6th Tech. Conf. Artificial Insemination Reprod. Natl. Assoc. Anim. Breeders, Columbia, MO, 1 10 pp. Anzar, M. and Graham, E.F. (1993). Animal Reprod. Sci. 31: Anzar, M. and Graham, E.F. (1995). Theriogenology 43: Anzar, M. and Graham, E.F. (1996). Theriogenology 45: Anzar, M. et al (1997). Theriogenology 47: 844 Bane, A. and Nicander, L. (1966). J. Reprod. Fert. 11: 133. Bhat, V. et al (2004). Indian J. Anim. Reprod. 25(1): Bhosrekar, M.R. et al (1991). Anim. Reprod. Sci. 26: Bhosrekar, M.R. et al (1992). Indian J. Anim. Sci. 62: Chauhan, S.S. et al (1993). Indian J. Anim. Sci. 63 (3): Chenoweth, P.J. et al (2003). Proceedings of the 16th Western Dairy Management Conference, March Clarke, I.J. and Tilbrook, A.J. (1992). Anim. Reprod. Sci. 28: Dixit, N.K. et al (1984). Theriogenology 24: El- Wishy, 1978; Younis, (1996) Malik et al., (1974) and Ahmad et al., (1984). Citated by Waltl, B.F. et al (2006).Anim. Reprod. Sci. 95: Foster, J. et al (1970). J. Anim. Sci. 30: Gahlot, P.S. and Kohli, T.S. (1981). Indian Vet. J. 58: Gahlot, P.S. et al (1990). Indian J. Anim. Reprod. 11(2): Goswami, S.C. et al (1991). Indian J. Anim. Sci. 61: Goyal, R. L. et al (1996). Theriogenology 46: Graham, E.F. and Graham, J.K. (1990). J. Dairy Sci. 73: Graham, E.F. et al (1976). VIII International Congress, Animal Reproduction Artificial Insemination, Cracow, IV: Hafs, H. D. (1972). In Proc. 4th Tech. Conf. Artificial Insemination Reprod. Natl. Assoc. Anim. Breeders, Columbia, MO, pp. Hallap, T. et al (2004). Theriogenology 62: Helbig, L. et al (2007). Anim. Reprod. Sci., 97: Igboeli. G. et al. Ainm. Reprod. Sci. 14: Januskauskas, A. et al (2005). Theriogenology, 63: Kammar, N.F. and Gangadhar, K.S. (1998). Indian J. of Anim. Reprod. 19(2): Kerruish, B.M. (1955). Br. J. Anim. Behav. 3: Khate, K. (2005). M.V.Sc.Tthesis in Animal Genetics and Breeding, NDRI, Karnal. Knop, K. et al (2005). Ainm. Reprod. Sci. 89: Kodagali, S.B. et al (1980). Indian J. Anim. Health, 19:31-34.

14 92 AGRICUTURAL REVIEWS Koonjaenak, S. et al (2007). Asian J. Androl. 9 (1): Kotayya, K., and Rao, A.V.H. (1981). Indian Vet. J. 88(7): Krishnan, R. et al (2003). National seminar on Frozen semen technology, Dhoni, Kerala, India, pp Kumar Ajeet et al (2003). Indian J. Anim. Sci. 73(8): Kumar, N. et al (1999). Indian J. Anim. Sci. 69: Larson, D. et al (1982). In Proc. 9th Tech. Conf. on Artificial Insemination and Reprod. Natl. Assoc. Anim. Breeders, Columbia, MO, Madan, M.L. (1985). Proc First World Buffalo Congress, Cairo. P Mandal, D.K. et al (2000). Indian J. Dairy Sci. 53: Manik, R.S. and Mudgal, V.D. (1984). World Review of Anim. Produc. 20 (4): 45-50, October- December: 1984 Mann, T. and Mann, C.L. (1981). Male Reproductive, Function and Semen. Springer- Verlag Berlin. Heidelberg, New York, USA. Mathevon, M. et al (1998). J. Dairy Sci. 1: Mathew, A. et al (1984). Anim Breed Abst 51(2): 856. Mathur, A.K. et al (2002). J. Livestock Poultry Prodn., 18(1-2): Mathur, A.K. et al (2003). Indian J. Anim. Sci. 73(8): Maurya, V.P. and Tuli, R.K. (2003). Asian- Aust. J. anim. Sci. 16 (10): Murugavel, K. et al (1997). Indian J. Anim. Sci. 67: Murugavel, K. et al (2000). Indian Vet. Med. J. 24: Mustafa, G. et al (1998). Theriogenology 50: Oloufa, M.M. et al (1959). Indian J. Dairy Sci. 12: Panghal, V.S. and Tuli, R.K. (1999). Indian J. Dairy sci. 52: Pankaj, P.K. (2006). Ph.D. Thesis, NDRI Deemed University, Karnal, Haryana. Parrish, J.J. et al (1995). Theriogenology 44: Prosser, J. J. (1968). In: Proc. 2nd Tech. Conf. Artificial Insemination Reprod. Natl. Assoc. Anim. Breeders, Columbia, MO, 44 45pp. Radhakrishnan, R. et al (1975). Indian Vet. J. 52: Rana, C.M. et al (2004). Indian J. Dairy Sci., 57(2): Rao, A.R. (1976). Indian Vet. J. 53: Rao, A.R. et al (1980). Theriogenology 14 (1): Rao, K.R. et al (1995). Indian Vet. J. 72: Rao, V.J. and Venkataswami, V. (1971). Indian Vet. J. 48: Ravimurgan, T. et al (2003). Indian Buffalo J., 1(2): Rekwot, P.I et al. Anim. Reprod. Sci. 14 : Samper, J.C. (1990). Ph.D. dissertation, University of Minnesota, St. Paul, MN; p Saxena, V.B., and Tripathi, S.S. (1976). Indian J. Anim. Sci. 46(1): Saxena, V.B., and Tripathi, S.S. (1977). Indian Vet. J. 54: Sethi, R.K. et al (1989). Indian Vet. J. 59: Shannon, P. (1972). Proc. 7th Int. Congr. Anim. Reprod. and A.I., Munich, Germany vol. 2 pp Singh, P. (2003). 4th Asian Buffalo Congr. On Buffalo for Food Security and Rural Employment, from 25-28th Feb., 2003, held in New Delhi. pp Singh, P. and Raina, V.S. (2000). Asian-Aust J. Anim. Sci., 13(7): Singh, R. et al (2003). Indian J. Anim. Sci. 73: Sudheer, S. and Xavier, C.J. (2000). Indian J. of Anim. Reprod. 21(1): Suryaprakasam, T.B. and Rao, A.V.N. (1993). Indian Vet. J. 70: Tomar, N.S. et al (1966). Indian J. Dairy Sci. 19: Tripathi, S.S., and Saxena, V.B. (1976). Indian J. Anim. Sci. 46: Tuli, R.K. and Singh, M. (1983). Theriogenology 20: Tyagi, S. et al (2006). Indian J. Anim. Reprod. 27(1): Ulgen G. et al (2004). Uludag Univ. J. Fac. Vet. Med. 23, 1-2-3: Venkatareddy, J. et al (1991). Indian J. of Anim. Produc. 12:

15 Vol. 30, No. 2, 2009 Venkataswami, V. et al (1972). Indian Vet. J. 49: Vyas, S. et al (1991). Indian J. Anim. Sci. 61(7): Vyas, S. et al (1992). Indian J Anim Sci 62: Yates, J.H. et al (2003). Theriogenology 60(9): Zafar, A.H. et al (1988). Buffalo J. 4(1):

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