Buffalo Bulletin (September 2009) Vol.28 No.3

STUDIES ON SOME BIOCHEMICAL CONSTITUENTS OF MURRAH BUFFALO(BUBALUS BUBALIS) SEMINAL PLASMA

M.K. Shukla1, A.K. Misra2 and H.P. Gupta2

1Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary Science and A.H.(JNKVV), Kuthulia, Rewa-486001, Madhya Pradesh, India2G.B. Pant University of Agriculture and Technology, Pantnagar, Udhamsinghnagar, Uttranchal-263145, India

ABSTRACT

The chemical composition of the seminalplasma of the three bulls, using 10 ejaculates fromeach bull, was analyzed to estimate concentrationof calcium, chloride, inorganic phosphorus,potassium, magnesium, sodium, bicarbonate, totalcholesterol, citric acid, and GOT. The respectiveaverages of the calcium, chloride, inorganicphosphorus, potassium, magnesium, sodium,bicarbonate, total cholesterol, citric acid, and GOTin the seminal plasma were 44.95 + 0.96 (mg %),366.73 + 53.69 (mg %), 6.82 + 1.63 (mg %), 98.18+ 11.67 (mg %), 6.61 + 0.49 (mg %), 106.46 + 11.92(mg %), 9.49 + 1.26 (milliequivalents per liter), 53.67+ 8.72 (mg %), 499.09 + 18.46 (mg %) and 87.45 +13.52 (Units ml-1). All the chemical constituents inthe three bulls’ semen were in the normal rangedescribed for the species. There was significantdifference (P<0.05) in calcium and magnesiumconcentration in seminal plasma of the three bulls,as the bull MB III had lower calcium content andMB II had higher magnesium content as comparedto the other two bulls.

Keywords: seminal plasma, semen, buffalo

INTRODUCTION

Composition of semen has been analyzedin many species including water buffalo for the studyof various biochemical and enzymatic constituents

of semen (Mann, 1964; Bhattacharya, 1974; Dabaset al., 1986; Dhami and Sahni, 1994; Kanwal et al.,1998; Prasad et al., 2000). Among the most importantaspects of the studies on spermatozoal metabolismhas been the understanding of the chemical pathwaysinvolved in energy utilization (White, 1958) andmaintenance of osmotic balance by ions present inthe semen (Hawk et al., 1964; Kanakraj andKrishnamurthy, 1984; Nath, 1988) which are animportant determinant of sperm viability. The presentstudy was conducted to study the biochemicalcomposition of the seminal plasma of Murrah bullsmaintained in Tarai region of Uttarakhand state ofIndia.

MATERIALS AND METHODS

Three Murrah buffalo bulls maintainedunder identical environmental and managementalconditions comprised the experimental animals forthe study. Ten semen samples from each bull werecollected in sterilized goat artificial vagina.Immediately after collection semen samples weresubjected to centrifugation at 1500 g for 30 minutesto obtain the seminal plasma, which wassubsequently separated and stored at -20oC tillfurther analysis. The estimation of the minerals inthe seminal plasma was done by the autoanalyser(NEXCT Random Access Bench Top Analyser,Schiapparelli Biosystem, USA).

The data were analyzed by analysis ofvariance (one way) using standard statistical methodsof Snedecor and Cochran, 1967.

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RESULTS AND DISCUSSION

The average values of the biochemicalconstituents are summarized in Table 1. The overallaverage of calcium (44.95 + 0.96 mg %) in thepresent study is comparable to the previous reports(40.0 + 2.0: Roy et al., 1960; 43.45 + 1.49: Singh etal., 1970; 41.11: Reddy and Raja, 1979) but werehigher than that reported by Kanwal et al., 1998(29.97 + 5.15). Higher calcium content in the semenis reported to have depressing effect on spermmetabolism (Mann and Mann, 1981). The ionspresent in the semen of bulls helps in stimulating themotility and glycolysis and counteract the depressingeffect of calcium present in the semen (Mann andMann, 1981).

The magnesium (6.61 + 0.49) content in theseminal plasma of the three Murrah bulls in thepresent study was higher than the findings of others(5.24 + 0.49: Singh et al., 1970; 5.91: Reddy andRaja, 1979), however, significantly higher (9.81 mg%) magnesium content has also been reported byCragle et al. (1958). Low levels of magnesium areassociated with comparatively lower motility of thesemen sample (Singh, 1970). The chloride content(366.73 + 53.69), in the present study, was also inagreement with the previous reports (373.55 + 55.0:Roy et al., 1960; 347.50: Singh et al., 1969) but wasconsiderably higher than 269.13 + 7.05, reported byDhami and Sahni, 1993. The bicarbonate content ofseminal plasma in the present study was also in tunewith the findings of Singh et al. (1969).

Different superscripts within a row indicate significant difference (P<0.05).

Table 1. Chemical composition (Mean + SE) of Murrah seminal plasma.

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Chemical constituent MB I (n=10)

MB II (n=10)

MB III (n=10)

Overall average (n= 30)

Calcium (mg/ 100 ml) 46.25a (+1.96)

47.30a (+1.38)

41.30b (+1.46)

44.95 (+0.96)

Chloride (mg/ 100 ml)

360.76a (+68.26)

371.53a (+75.38)

367.91a (+63.35)

366.73 (+53.69)

Inorganic Phosphorus (mg/ 100 ml)

7.03a (+1.36)

6.93a (+1.20)

6.51a (+1.71)

6.82 (+1.63)

Potassium (mg/ 100 ml)

98.07a (+12.36)

97.39a (+11.93)

99.07a (+12.93)

98.18 (+11.67)

Magnesium (mg/ 100 ml)

5.62a (+0.37)

8.05b(+0.31)

6.15a (+0.47)

6.61 (+0.49)

Sodium (mg/ 100 ml)

109.31a (+12.80)

103.09a (+12.07)

106.97a (+12.91)

106.46 (+11.92)

Bicarbonate (Milliequivalents per liter)

9.37a (+1.33)

9.99a (+1.91)

9.11a (+1.95)

9.49 (+1.26)

Total Cholesterol (mg/ 100 ml)

54.91a (+9.20)

52.36a (+9.13)

53.73a (+8.63)

53.67 (+8.72)

Citric acid (mg/ 100 ml)

491.69a (+21.47)

501.23a (+27.39)

504.36a (+19.70)

499.09 (+18.46)

Glutamic Oxalo-acetic transaminase (unit ml-1)

87.09a (+13.71)

89.91a (+16.47)

85.36a (+14.61)

87.45 (+13.52)

Buffalo Bulletin (September 2009) Vol.28 No.3

Sodium is an extracellular element whilePotassium is intracellular in nature, and there is anopinion that normal ionic equilibrium and osmoticpressure are maintained by these ions (Hawk et al.,1964). A positive correlation of sodium and potassiumcontent in the semen is reported to be responsiblefor the maintenance of osmolarity and metabolicactivity of the spermatozoa (Nath, 1988). Theaverage potassium content (98.18 + 11.67) in thepresent experiments was comparable to the findingsof the earlier researchers (101.60 + 4.45: Singh etal., 1970; 98.04 + 2.77: Kanakraj and Krishnamurthy,1984; 103.56 + 3.15: Dhami and Sahni, 1993)whereas the sodium (106.46 + 11.92) content waslower than the previous reports (> 250.0: Singh etal., 1969; 186.89 + 10.90: Singh et al., 1970; 139.00+ 3.00: Gupta and Tripathi, 1983). The potassiumcontent was also found to be negatively correlatedwith sperm abnormalities, suggesting that the highpotassium content in the seminal milieu might beresponsible for maintenance of spermatozoalconfiguration (Nath, 1988). Potassium concentrationhas also been reported to be significantly correlatedwith initial sperm motility (-0.16) and spermconcentration (0.18) whereas a significant correlationof the sodium concentration with the live spermpercent has been observed (Nath, 1988). Thecontent of potassium and sodium in the seminalplasma in the present study indicates fairly goodsemen quality of the three bulls.

The concentration of inorganic phosphorus(6.82 + 1.63) in the seminal plasma of the three bullsin the current experiments was in conformation withthe previous reports (6.4 + 0.6: Roy et al., 1960;7.56 + 0.42: Singh et al., 1970) whereas an evenhigher concentration (12.28 + 0.37: Dhami and Sahni,1993) has been reported. The phospholipid is requiredfor the preservation of the sperm membrane integrity(Singh, 1969).

The citric acid content in the semen is knownto be influenced by the circulatory level of androgen(Dondero et al., 1972). The citric acid (499.09 +18.46) content of seminal plasma in the currentinvestigation was in fair agreement with the findingsof Singh et al., (1970). The fairly normal levels ofcitric acid in the present investigations indicate thebetter buffering capacity of the seminal plasma,which could have helped in maintaining the sperm

abnormalities within the limits as citric acid is knownto help in maintaining morphological integrity of thespermatozoa (Kumar, 1986).

Cholesterol, the precursor of androgen(Singh, 1969), is one of the main constituents of theseminal plasma that has a decisive cryoprotectiveeffect (Rattan et al., 1973). The average totalcholesterol level (53.67 + 8.72 mg %) in the presentinvestigation is possibly suggestive of the normaltestosterone levels to maintain desired sex behaviourand libido of the three bulls. The total cholesterolcontent of Murrah seminal plasma in the presentexperiment is comparable to the findings of theprevious researchers (55.02 + 17.46: Prabhu et al.,1973; 53.00 + 8.88: Varshney et al., 1978).

Poor storage of buffalo semen may be dueto the higher leakage of enzymes in the seminalplasma (Chuahan and Srivastava, 1973) as theincreased enzyme leakage is reported to bear anegative correlation with sperm motility, live spermper cent and acrosome damage (Sharma et al.,2001). In the present investigation, glutamic oxalo-transaminase (GOT) content in the Murrah seminalplasma was 87.45 + 13.52 units /ml, which is in tunewith the previous reports of Varshney et al., 1978(83.50 + 3.00) and Kumar, 1986 (95.88 + 15.32).GOT has been reported to be directly correlatedwith the sperm concentration (Saxena et al., 1978)and acrosomal damage (Sharma et al., 2001).

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