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		  <p class="Type-of-Article"><span class="CharOverride-1">Research Article</span></p>
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			<p class="title- ParaOverride-1">&nbsp;</p>
			<p class="title- ParaOverride-1">Biochemical Profiles in Freezable and Non-Freezable Ejaculates of Mithun (Bos frontalis) Semen</p>
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			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-3">Ponraj Perumal</span><span class="CharOverride-4">1</span><span class="CharOverride-3">, Suresh Kumar Srivastava</span><span class="CharOverride-4">2</span><span class="CharOverride-3">, Kezhavituo Vupru</span><span class="CharOverride-4">3</span><span class="CharOverride-3">, Kobu Khate</span><span class="CharOverride-4">3</span><span class="CharOverride-3">, Muthusamy Veeraselvam</span><span class="CharOverride-4">4</span><span class="CharOverride-3">, Chandan Rajkhowa</span><span class="CharOverride-4">5</span></p>
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			<p class="Affiliations ParaOverride-1">1Animal Reproduction Laboratory, ICAR- National Research Centre on Mithun, Jharnapani, Nagaland – 797 106 (India); 2 Animal Reproduction Division, ICAR- Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh – 243 122; 3 Farm Manager, ICAR- National Research Centre on Mithun, Jharnapani, Nagaland – 797 106, India; 4Veterinary University Training and Research Centre, TANUVAS, Salem, India; 5 ICAR- National Research Centre on Mithun, Jharnapani, Nagaland – 797 106 (India).</p>
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			<p class="Abstract ParaOverride-1">&nbsp;</p>
		  <p class="Abstract ParaOverride-1"><span class="CharOverride-5">Abstract</span> | <span lang="en-US">The present study was undertaken to measure the biochemical profiles of freezable and non- freezable ejaculates of Mithun. Fifty ejaculates (twenty five ejaculates each from freezable and non- freezable ones) were collected from matured Mithun bulls. Biochemical profiles viz., alkaline phosphatase (ALP), acid phosphatase (ACP), </span><span class="Emphasis CharOverride-6" lang="en-US">aspartate aminotransferase (AST), alanine aminotransferase (ALT), </span><span lang="en-US">lactic acid dehydrogenase (LDH), calcium (Ca), magnesium (Mg), zinc (Zn), chloride (Cl), citric acid (CA), total seminal plasma protein (TSPP), inorganic phosphorous (IP), fructose and cholesterol concentration in sperm were estimated. The result revealed that these parameters varied significantly (P&lt; 0.05) between the freezable and non-freezable ejaculates. Freezable ejaculates has significantly (P&lt; 0.05) higher ALP, ACP, LDH, Ca, Mg, Zn, citric acid, TSPP, IP, fructose and cholesterol concentration and significantly (P&lt; 0.05) lower Cl concentration than the non- freezable ejaculates. There was a correlation among the biochemicals such as ALP, ACP, LDH, Ca, Mg, Zn, CA, TSPP, IP, fructose, cholesterol and these biochemicals were negatively correlated with AST, ALT and Cl in both freezable and non-freezable ejaculates. It was concluded that most of the biochemical parameters were higher in freezable ejaculates than the non-freezable ejaculates, indicates that freezable ejaculates have has structural stability than the non-freezable ones that leads to freezable sperm has </span><span class="annotation-reference CharOverride-7" lang="en-US">h</span><span lang="en-US">igher functional structures to move faster and in forward direction.</span></p>
			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-5">Keywords </span>| <span lang="en-US">Mithun, Biochemical profiles, Freezable and non-freezable ejaculates </span></p>
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			<p class="Editor----Citation">&nbsp;</p>
			<p class="Editor----Citation"><span class="CharOverride-9">Editor</span> | Kuldeep Dhama, Indian Veterinary Research Institute, Uttar Pradesh, India.</p>
            
			<p class="Editor----Citation"><span class="CharOverride-5">Received</span> | October 31, 2014; <span class="CharOverride-5">Revised</span> | November 29, 2014; <span class="CharOverride-5">Accepted</span> | November 30, 2014; <span class="CharOverride-5">Published</span> | December 15, 2014&#9;&#9;</p>
			
          <p class="Editor----Citation"><span class="CharOverride-5">*Correspondence</span> | Ponraj Perumal, National Research Centre on Mithun, Jharnapani, Nagaland, India; <span class="CharOverride-5">Email:</span> perumalponraj@gmail.com</p>
			
          <p class="Editor----Citation"><span class="CharOverride-5">Citation</span> | Perumal P, Srivastava SK, Vupru K, Khate K, Veeraselvam M, Rajkhowa C (2015). Biochemical profiles in freezable and non-freezable ejaculates of Mithun (Bos frontalis) semen. Adv. Anim. Vet. Sci. 3(1): 19-26.  </p>
			<p class="Editor----Citation"><span class="CharOverride-9">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.aavs/2015/3.1.19.26"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.aavs/2015/3.1.19.26</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-5" lang="en-US">ISSN (Online)</span> | 2307-8316; 
          <span class="Editor---Citation CharOverride-5" lang="en-US">ISSN (Print)  | </span>2309-3331</p>
            
		  <p class="Editor----Citation"><span class="CharOverride-9">Copyright </span>© 2015 Perumal et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</p>
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			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">INTRODUCTION </p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Caps-on-First-Para ParaOverride-1"><span class="_idGenDropcap-1">M</span>ithun (<span class="CharOverride-11">Bos frontalis</span>) is a semi-wild, free–range bovine species present in the North-Eastern Hill (NEH) region of India and is believed to have originated more than 8000 years ago from wild Indian gaur (<span class="CharOverride-11">Bos gaurus) </span>(<a href="#Simoons-FJ--1984-"><span class="Hyperlink">Simoons, 1984</span></a>). It occupies an important place in the socio-cultural and economic life of the tribal population especially in Arunachal Pradesh, Nagaland, Manipur and Mizoram states of India. Mithun <span class="annotation-reference">p</span>opulation is decreasing (Livestock census of India, 2007) due to various reasons such as insufficient number of breeding bulls, intensive inbreeding and lack of proper breeding management practices. Dwindling population of Mithun is a cause of concern which requires greater effort from all quarters to conserve the species bearing much effect on the socio-economic status of tribes. In Mithun natural breeding is being followed <span class="annotation-reference">w</span>ith accompanied limitations such as cost and disease transmission. Under such circumstances, wider application of artificial breeding of Mithun appears a viable option for conserving the species. </p>
		  <p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The seminal plasma is a highly complex biological fluid, secreted from various accessory sex glands and testes and contains various biochemical constituents such as cholesterol, sugars, proteins, metabolic, intra cellular and antioxidant enzymes, mineral elements (<a href="#Kulkarni-BA--Dhande-SG--Patankar-DD--1996-."><span class="Hyperlink">Kulkarni et al., 1996</span></a>). The constituents of seminal plasma have various functions such acquisition of motility, capacitation, acrosome reaction and fertilizing capacity of spermatozoa (<a href="#Kulkarni-BA--Dhande-SG--Patankar-DD--1996-."><span class="Hyperlink">Kulkarni et al., 1996</span></a>). Several studies were conducted on composition of seminal plasma in different domestic animal species (<a href="#Mann-T--1964-."><span class="Hyperlink">Mann, 1964</span></a>; <a href="#Dabas-YPS--Verma-MC--Tripathi-SS--Saxena-VB--1984"><span class="Hyperlink">Dabas&#160;et al., 1984</span></a>; <a href="#Dhami-AJ--Sahni-KL--1994-."><span class="Hyperlink">Dhami and Sahni, 1994</span></a>). Further, perusal of literatures revealed meagre information on the composition of seminal plasma in Mithun Bulls. Therefore, the study was designed to measure the biochemical composition of seminal plasma in Mithun to aid in for future semen preservation.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">MATERIALS AND METHODS</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Ten apparently healthy Mithun Bulls of 3 to 5yr of age were selected from the herd in Mithun farm, NRC on Mithun, Jharnapani, Nagaland. The average body weight was 501 kg (493 to 507 kg) with good body condition (score 5-6) maintained under uniform feeding (farm schedule) and managemental conditions. A total of 50 ejaculates were collected through rectal massage method. These ejaculates were splited into freezable and non-freezable ejaculates based on the post thaw motility (<a href="#Rao-AVN--Rao-YVR--1996-."><span class="Hyperlink">Rao and Rao, 1996</span></a>; <a href="#Prasad-JK--Kumar-S--Mohan-G--Shankar-U--Agarwal-SK--2000-."><span class="Hyperlink">Prasad et al., 2000</span></a>). Ejaculates having post thaw motility 40% and above were considered as freezable ejaculates whereas non-freezable ejaculates were considered with a post thaw motility less than 40%. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The biochemical profiles <span class="CharOverride-11">viz</span>., <span class="annotation-reference CharOverride-14">A</span>LP, ACP, AST, ALT, LDH activity, Ca, Mg, Zn, Na, K, inorganic phosphorous, chloride, fructose, total seminal plasma protein and total cholesterol of the semen were estimated by commercial available kits (Cayman chemical company, USA). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The results were analysed statistically between the freezable and non-freezable ejaculates with the student ‘t’ test using the SPSS (version 15.0; SPSS, Chicago, IL) and expressed as the mean ± S.E.M. Significant difference values of (p&lt;0.05) were considered to be statistically significant after arcsine transformation of percentage data. Correlation among the biochemical parameters was estimated with Pearson’s correlation coefficient. Differences at (p &lt; 0.05) were considered to be statistically significant.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">RESULTS AND DISCUSSION </p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">In the present study, the result revealed that estimated different biochemical profiles varied significantly (P&lt;0.05) between the freezable and non-freezable ejaculates (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). Freezable ejaculates has significantly (P&lt; 0.05) higher ALP, ACP, LDH, Ca, Mg, Zn, citric acid, TSPP, IP, fructose and cholesterol and significantly lower <span class="annotation-reference CharOverride-14">C</span>l content than non- freezable ejaculates. There was positive correlation among the biochemicals such as ALP, ACP, LDH, Ca, Mg, Zn, CA, TSPP, IP, fructose, cholesterol and these biochemicals were negatively correlated with AST, ALT and Cl in both freezable (<a href="#Table-2"><span class="Hyperlink">Table 2</span></a>) and non-freezable (<a href="#Table-3"><span class="Hyperlink">Table 3</span></a>) ejaculates. The present information may aid in getting good quality semen for artificial insemination by selecting freezable ejaculates based on biochemical constituents of seminal plasma.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">Phosphatase enzymes such as ALP and ACP levels in seminal plasma are very important for sperm metabolism as well as sperm functions (<a href="#Brooks-DE--1990-"><span class="Hyperlink">Brooks, 1990</span></a>). Therefore, estimates of these enzymes have been recommended as biomarkers for assessment of semen quality (<a href="#Pesch-S--Bergmann-M--Bostedt-H--2006"><span class="Hyperlink">Pesch et al., 2006</span></a>). ALP in seminal plasma is primarily of testicular and epididymal origin and can be used as a clinical ejaculatory marker to differentiate azoospermia or oligospermia from ejaculatory failure (<a href="#Turner-RM--McDonnell-SM--2003-"><span class="Hyperlink">Turner and McDonnell, 2003</span></a>). In the present study, the ALP was significantly decreased in the non- freezable ejaculates than the freezable ejaculates (<a href="#Pesch-S--Bergmann-M--Bostedt-H--2006"><span class="Hyperlink">Pesch et al., 2006</span></a>). ACP is especially localized in corpus epididymidis, ductus epididymidis and vas deferens, but it is thought to be <span class="Body-Text ParaOverride-1">an indicator for thesecretory function of prostate (<a href="#Ciereszko-A--Glogowski-J--Strzezek-J--Demianowicz-W--1992"><span class="Hyperlink">Ciereszko et al., 1992</span></a>).</span></p>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-2"><span class="CharOverride-15"><a id="Table-1-"></a>Table 1:</span><span class="CharOverride-16"> Mean (±S.E.) biochemical attributes of freezable and non- freezable Mithun semen</span><span class="Title CharOverride-17" lang="en-US"><a id="x.29713"></a></span></p>
			<table width="657" height="455" class="Table-Style-1 TableOverride-1" id="table-1">
				<colgroup>
					<col class="_idGenTableRowColumn-1" />
					<col class="_idGenTableRowColumn-2" />
					<col class="_idGenTableRowColumn-3" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">Biochemical attributes</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">Freezable semen (n=25) </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">Non- freezable semen (n=25) </span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALP (KAU/100 mL)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">230.66 ± 3.40</span><span class="Title CharOverride-18">a</span><span class="Title">                    </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">180.70 ± 5.22</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ACP (KAU/100 mL)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">246.75 ± 3.75</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">189.86 ± 6.17</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">AST (µ mole/litre)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">78.62 ± 1.64</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">89.08 ± 3.18</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALT (µ mole/litre)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">13.19 ± 1.30</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">17.67 ± 1.37</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">LDH (IU/Litre)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">317.99 ± 3.58</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">244.11 ± 6.53</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Ca (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">35.13 ± 2.14</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">25.41 ± 1.72</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Mg (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">7.27 ± 0.83</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">5.41 ± 1.14</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Zn (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">3.80 ± 0.66</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">2.46 ± 0.90</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Chloride (mmol/L)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">320.35 ± 5.17</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">402.94 ± 7.25</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Citric Acid (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">552.18 ± 6.78</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">443.73 ± 7.13</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Total Protein (g/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">8.22 ± 0.86</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">6.18 ± 0.78</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Inorganic Phosphorous (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">9.20 ± 1.12</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">6.91 ± 0.83</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Fructose (mg/dl)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">508.41 ± 6.28</span><span class="Title CharOverride-18">a</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">424.85 ± 5.89</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Total Cholesterol (µg/10</span><span class="Title CharOverride-18">8</span><span class="Title">sperm)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">28.55 ± 1.45</span><span class="Title CharOverride-18">a</span><span class="Title"> </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">22.71 ± 1.44</span><span class="Title CharOverride-18">b</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-13">Means with different superscript within rows differ significantly (</span><span class="CharOverride-19">P </span><span class="CharOverride-13">&lt; 0.05); n= Number of ejaculates</span></p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Like phosphatse, AST and ALT are essential for metabolic processes, which provide energy for motility, viability and fertilizing ability of spermatozoa and these transaminase activities in the ejaculates are good indicators of semen quality as these enzymes measure stability of sperm membrane (<a href="#Lopez-ML--Grez-P--Gribbel-I--Bustos-Obreg-n-E--1989"><span class="Hyperlink">Lopez et al., 1989</span></a>). Increasing abnormal sperm percentage in the ejaculate causes increased concentration of these transaminase enzymes in the extra cellular fluid which might be due to damage of sperm plasma membrane and leakage of enzymes from spermatozoa (<a href="#Dogan-I--Polat-U--Nur-Z--2009-"><span class="Hyperlink">Dogan et al., 2009</span></a>). Moreover, increase in AST and ALT activities of seminal plasma of non- freezable ejaculates could be due to structural instability of the sperm or fragile nature of sperm membrane (<a href="#Corteel-JM--1980-."><span class="Hyperlink">Corteel, 1980</span></a>) and less intactness of membrane of acrosome, plasma, mitochondria and flagella of the sperm. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">LDH is an essential enzyme of almost universal distribution in the body which catalyses the reversible transamination of pyruvate to lactate. In semen, it is chiefly located in the mid-piece region (<a href="#Dhami-AJ--Sahni-KL--1994-."><span class="Hyperlink">Dhami and Sahni, 1994</span></a>). Like transaminases, LDH is also an essential enzyme responsible for metabolic processes, which provide energy for viability, motility, capacitation and fertilizing ability of spermatozoa (<a href="#Sirat-MP--Sinha-AK--Singh-BK--Prasad-L--1996"><span class="Hyperlink">Sirat et al., 1996</span></a>). It has been proposed that the seminal fluid LDH can be used as good indicator of sperm viability and membrane stability (<a href="#Stamatiads-K--Karayanidis-A--Tsakalof-P--1984-."><span class="Hyperlink">Stamatiads et al., 1984</span></a>).<span class="apple-converted-space CharOverride-21">&#160; </span><a href="#Pesch-S--Bergmann-M--Bostedt-H--2006"><span class="Hyperlink">Pesch et al. (2006)</span></a> reported that there was positive correlation among LDH activities, total sperm motility, progressive motility and liveability of sperm in fresh ejaculates, which may indicates that extracellular LDH ensures metabolism of spermatozoa. In the present study, extracellular LDH concentration was increased significantly (p&lt; 0.05) in seminal plasma of freezable ejaculates (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>) than the non-freezable ejaculates. It has thus been proposed that higher LDH levels in seminal plasma of fresh semen can be used as a good indicator of higher motility, progressive motility and living sperm of ejaculates (<a href="#Dube-GD--Dwaraknath-PK--Vyas-KK--1982"><span class="Hyperlink">Dube et al., 1982</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The Ca, Mg, Zn and IP of ejaculate were significantly higher in freezable ejaculates than the non- freezable ejaculates, whereas chloride content was higher in non-freezable ejaculates in the present study (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). Zn plays an essential role in normal testicular development, spermatogenesis and sperm motility (<a href="#Wong-WY--Flik-G--Groenen-PM--Swinkels...2001"><span class="Hyperlink">Wong et al., 2001</span></a>). It is also a cofactor for a number of metalloenzymes in animals. Zn plays an important role in prostate, epididymal and testicular functions (<a href="#Ebisch-TMW--Van-Heerde-WL--Thomos-CMG--Vander-Put-N--Wong-WY--Steegers-Theunissen-RPM--2003"><span class="Hyperlink">Ebisch et al., 2003</span></a>). Zn has been reported to influence the process of spermatogenesis (<a href="#Wong-WY--Merkus-HM--Thomas-CM--Menkveld-R--Zielthuis-GA--Steegers--Theunissen-RP--2002-."><span class="Hyperlink">Wong et al., 2002</span></a>), controls sperm motility (<a href="#Wroblewski-N--Schill-WB--Henkel-R--2003-."><span class="Hyperlink">Wroblewski et al., 2003</span></a>), stability of sperm membrane (<a href="#Kendall-NR--McMullan-S--Green-A--Rodway-RG--2000"><span class="Hyperlink">Kendall et al., 2000</span></a>), preserves the ability of sperm nuclear chromatin to undergo de-condensation and modulates sperm functions (<a href="#Suruki-T--Nakajima-K--Yamamoto-A--Yamanaka-H--1995-"><span class="Hyperlink">Suruki</span><span class="Hyperlink"> et al., 1995</span></a>). Hypozinkemia leads to gonad dysfunction, decreased testicular weight, atrophy of seminiferous tubules and complete cessation of spermatogenesis (<a href="#Martin-GB--White-CL--Markey-CM--Blackberry-MA--1994"><span class="Hyperlink">Martin et al., 1994</span></a>). </p>
			<p>&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Table-2"></a>Table 2:</span> Correlation coefficient among the biochemical attributes of Freezable ejaculates in Mithun (<span class="CharOverride-11">Bos frontalis</span>) bulls</p>
			<table width="657" class="Table-Style-1 TableOverride-1" id="table-2">
				<colgroup>
					<col class="_idGenTableRowColumn-6" />
					<col class="_idGenTableRowColumn-7" />
					<col class="_idGenTableRowColumn-8" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-10" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-11" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-9" />
					<col class="_idGenTableRowColumn-10" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5"><a id="x.29909"></a>No</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">Parameters </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">5</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">11</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">12</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">13</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">14</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALP</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.88*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.46</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.76*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.77*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.61</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.46</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.82*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.85*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.79*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ACP</span></p>
						</td>
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.16</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.43</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.77*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.79*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.48</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.82*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.81*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.80*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.90*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.57</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">AST</span></p>
						</td>
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.37</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.68*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.46</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.23</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.34</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.11</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.90*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.51</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.55</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.68*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALT</span></p>
						</td>
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.47</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.67*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.79*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.80*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.51</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.91*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.81*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.76*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.57</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">5</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">LDH</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.59</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.44</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.55</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.24</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.81*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.74*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.78*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.50</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Ca</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.65*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.75*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.33</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.71*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.75*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.64*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Mg</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.48</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.68*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.63*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.80*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.43</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Zn</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.53</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.83*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.58</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Cl</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.41</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.42</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.62*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.16</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">CA </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.97*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.93*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">11</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">TP </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.91*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.83*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.79*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">12</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">IP</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.80*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.83*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">13</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">FR</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.51</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">14</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">CH </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1">ALP: alkaline phosphatase, ACP: acid phosphatse, AST: <span class="Emphasis CharOverride-16">aspartate aminotransferase, ALT: alanine aminotransferase, LDH: </span>lactic acid dehydrogenase, Ca: calcium, Mg: magnesium, Zn: zinc, Cl: chloride, CA: citric acid, TP: total protein, IP: inorganic phosphate, FR: fructose, CH: cholesterol in sperm, * Correlation coefficient were significant, p&lt; 0.05</p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Table-3"></a>Table 3:</span> Correlation coefficient among the biochemical attributes of non- freezable ejaculates in Mithun (<span class="CharOverride-11">Bos frontalis</span>) bulls</p>
			<table width="657" height="466" class="Table-Style-1 TableOverride-1" id="table-3">
				<colgroup>
					<col class="_idGenTableRowColumn-16" />
					<col class="_idGenTableRowColumn-17" />
					<col class="_idGenTableRowColumn-18" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-20" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-19" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-21">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5"><a id="x.30371"></a>No</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">Parameters </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">5</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">11</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">12</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">13</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title CharOverride-5">14</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-22">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALP</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.98*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.82*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.91*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.79*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.84*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.92*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.82*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ACP</span></p>
						</td>
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.75*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.92*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.97*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.85*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.75*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">AST</span></p>
						</td>
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.70*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.88*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.83*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.71*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.74*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.67*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.75*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.67*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.86*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">ALT</span></p>
						</td>
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.63*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.92*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.92*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.85*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.82*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.66*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">5</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">LDH</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.84*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.91*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.84*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Ca</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.69*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.59</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.71*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.81*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.65*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.91*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Mg</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.67*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.87*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.76*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Zn</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.98*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.92*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.85*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.75*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">Cl</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.94*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.88*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.82*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.95*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">-0.67*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">CA </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.96*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.98*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.93*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.84*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">11</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">TP </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.89*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.91*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.86*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">12</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">IP</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.828</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.85*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-23">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">13</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">FR</span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">0.72*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-12">
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">14</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">CH </span></p>
						</td>
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td />
						<td>
							<p class="Body-Text ParaOverride-2"><span class="Title">1.00</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1">ALP: alkaline phosphatase, ACP: acid phosphatse, AST: <span class="Emphasis CharOverride-16">aspartate aminotransferase, ALT: alanine aminotransferase, LDH: </span>lactic acid dehydrogenase, Ca: calcium, Mg: magnesium, Zn: zinc, Cl: chloride, CA: citric acid, TP: total protein, IP: inorganic phosphate, FR: fructose, CH: cholesterol in sperm, * Correlation coefficient were significant, p&lt; 0.05</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Calcium and Mg are required in many physiological processes as a regulator in all living cells, including spermatozoa. The presence of Mg <span class="CharOverride-18">2+</span> and Ca <span class="CharOverride-18">2+</span> ions is necessary for the last stage of capacitation <span class="annotation-reference CharOverride-14">f</span>ollowing acrosome reaction and hyperactive motility of spermatozoa. Ca flux control through the spermatozoal membrane is essential for fertilization (<a href="#Bailey-JL--Buhr-MM--1993-."><span class="Hyperlink">Bailey and Buhr, 1993</span></a>) and it was noticed that the inclusion of Ca<span class="CharOverride-18">2+</span> with calcimine to isolated ram caudal spermatozoa caused stimulation of flagellar beat activity (<a href="#Bradley-MP--Forrester-IT--1982-."><span class="Hyperlink">Bradley and Forrester, 1982</span></a>). But negative correlation between Ca concentration in seminal plasma and motility of bovine spermatozoa was also reported (<a href="#Machal-L--Chladek-G--Strakova-E--2002-."><span class="Hyperlink">Machal et al., 2002</span></a>). In the present study, concentration of Ca and Mg were significantly higher in freezable than the non- freezable ejaculates. <span class="annotation-reference CharOverride-14">C</span>alcium is positively correlated with sperm motility, viability and integrity of plasma and acrosomal membrane as Ca stabilizes the plasma membrane and influences its permeability and excitability. The higher correlation coefficient between seminal plasma Ca concentration and protein suggests that spermatozoon proteins as well as those of seminal plasma participate in extracellular Ca flow activation (<a href="#Marques-VA--Goulart-LR--Silva-AEDF--2000"><span class="Hyperlink">Marques et al., 2000</span></a>). Further, Ca is also required to stimulate the steriodogenesis in Leydig cells of the testis. Deficiency of Ca in the testes or accessory gland will leads to adverse effects on the normal function of the reproductive system and spermatogenesis (<a href="#Kaplan-A--Jack-R--Opheim-KE--Toivola-B--Lyon-AW--1995-"><span class="Hyperlink">Kaplan et al., 1995</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp; </p>
			<p class="Body-Text ParaOverride-1">   </p>
			<p class="Body-Text ParaOverride-1">Chloride levels were significantly (p&lt; 0.05) higher in the non- freezable ejaculates than the freezable ejaculates (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). The excess chloride could be toxic to bovine sperm which is neutralised by increased sodium levels (<a href="#Dhami-AJ--Sahni-KL--1994-."><span class="Hyperlink">Dhami and Sahni, 1994</span></a>). This could explain the reason for increased chloride in non- freezable ejaculates. In non-freezable ejaculates, seminal plasma fructose and metabolic enzyme concentrations are also low whereas chloride concentration is high (<a href="#Hirsch-IH--Jeyendran-RS--Sedor-J--Rosecrans-RR--Staas-WE--1991-"><span class="Hyperlink">Hirsch et al., 1991</span></a>). The low protein and high chloride content of the secretion is rather hostile to sperm. It is known that modulation of a variety of ion channels (like Cl) of spermatozoa is a characteristic event associated with capacitation and acrosome reaction of mammalian spermatozoa (<a href="#Barrier-Battut-I--Delajarraud-H--Legrand-E--Buyas-JF--Fieni-F--Tainturier-D--Thorin-C--Pouliquen-H--"><span class="Hyperlink">Barrier-Battut et al., 2002</span></a>). Hence, increasing the Cl level in seminal plasma may play a role in infertility. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">TSPP in the seminal plasma fluid influence various functions of the sperm such as capacitation, acrosome reaction, motility, DNA integrity and fertilization of the oocyte (<a href="#Moura-AA--Chapman-DA--Koc-H--Killian-GC--2007"><span class="Hyperlink">Moura et al., 2007</span></a>). The level of TSPP in fresh semen of freezable ejaculates was significantly higher than the non- freezable ejaculates (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). But some specific proteins might be responsible for the quality and freezability of semen which could not be differentiated between the two groups simply by estimating the total seminal plasma protein. Moreover, no report has so far been available on total seminal plasma protein level of freezable and non-freezable ejaculates in Mithun bull. <a href="#Dhami-AJ--Sahni-KL--1994-."><span class="Hyperlink">Dhami and Sahni (1994)</span></a> reported low mean protein value of static ejaculate than the motile ejaculate which supports the higher numerical value of TSPP in the highly motile samples in the present study. Similarly, <a href="#Singh-DM--Pangawkar-GR--Hundal-RS--Chaudhary-KC--1989-"><span class="Hyperlink">Singh et al. (1989)</span></a> reported a positive association of the protein values in the semen with its freezability. The reason could be the great importance of protein for motility and survival of spermatozoa during storage (<a href="#Singh-DM--Pangawkar-GR--Hundal-RS--Chaudhary-KC--1989-"><span class="Hyperlink">Singh et al., 1989</span></a>; <a href="#Moura-AA--Chapman-DA--Koc-H--Killian-GC--2007"><span class="Hyperlink">Moura et al., 2007</span></a>). Seminal plasm proteins are also known to have protective action towards sperm against lipid peroxidation (<a href="#Schonek-C--Braun-J--Einspanier-R--1996-"><span class="Hyperlink">Schonek et al., 1996</span></a>). But <a href="#Mohanty-DN--1999-."><span class="Hyperlink">Mohanty (1999)</span></a> reported higher TSPP in poor freezable bulls than the good freezable bulls. This variation could be due to different selection criteria of freezable and non- freezable ejaculates, frequency of collection, method employed for estimation, age and number of bulls studied and season (<a href="#Dhami-AJ--Sahni-KL--1994-."><span class="Hyperlink">Dhami and Sahni, 1994</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Fructose and citric acid are reported to play important roles in sperm motility and concentration, particularly with regard to energy metabolism, through glucose utilization (<a href="#Videla-E--Blanco-AM--Galli-ME--Fernandez-Collazo-E--1981-"><span class="Hyperlink">Videla et al., 1981</span></a>). Fructose is one of the major energy yielding nutritive substrates present in seminal fluid (<a href="#Gonzales-GF--Villena-A--1997-."><span class="Hyperlink">Gonzales et al., 1997</span></a>). It is secreted from the seminal vesicles and the accessory sex glands. Fructose is the major carbohydrate found in seminal plasma, provides over half the spermatozoa carbohydrate consumption and appears essential for normal sperm motility. In the present study, the fructose and citric acid concentration were reduced significantly in non- freezable ejaculates (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>) as indicates that the functions of accessory sex glands was affected as seminal fructose is positively correlated with semen volume and sperm motility (<a href="#Saeed-S--Khan-FA--Rehman-SB--Khan-DA--Ahmad-M--1994-"><span class="Hyperlink">Saeed et al., 1994</span></a>). The determination of fructose itself is of particular significant because there is a direct relationship between the fructose level in seminal plasma and the testosterone function of the interstitial cells of Leydig. Fructose values which fall below normal may be a consequence of inflammatory condition in the prostrate or seminal vesicles or structural abnormality of the seminal vesicles and their ducts (<a href="#Schirren-C--1983"><span class="Hyperlink">Schirren, 1983</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">It was concluded from the study that most of the biochemical parameters were significantly higher in freezable ejaculates in comparison to the non-freezable ejaculates of Mithun. This indicates that freezable sperm has structural stability than the non-freezable sperm that leads to freezable sperm has higher functional sperm structures to move faster and forward direction.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
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