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Residual Effects of Enrofloxacin Administration on Biochemical Attributes of the Semen of Barbari Bucks

Chandrima Sinha Brijesh Yadav Sarvajeet Yadav K. D. Singh
Vol 1(1), 30-36
DOI- http://dx.doi.org/10.5455/ijlr.20120204083514

The study was conducted to investigate the effect of enrofloxacin administration on aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AKP) and acid phosphatase (ACP) in seminal plasma of Barbari bucks. The bucks were administered enrofloxacin at the dose of 5mg/kg body weight intra-muscularly daily for 7 days. From each buck biweekly eighteen ejaculates (1st to 18th) were collected and one sample (0th) from each buck was collected before administration of enrofloxacin buck using a non-oestrous doe as a dummy. Seminal plasma was obtained after centrifuging semen and AST, ALT, ACP and AKP level was estimated besides protein concentration. AST and ALT levels were significantly higher (P≤0.01) and maximum in seminal plasma of 9th and 5th ejaculate respectively, as compared to ejaculate at 0 day. The AKP and ACP level increased significantly (P≤0.01) and was the maximum in the seminal plasma of 1st and 4th ejaculate respectively, as compared to ejaculate at 0 day. It may be concluded from this study that only after nine weeks of enrofolaxacin administration, the biochemical status of seminal plasma with respect to ALT, AST, AKP and ACP becomes normal which may be attributed to residual effect of enrofloxacin.


Keywords : Enrofloxacin Seminal Plasma Biochemistry

Introduction

The animals with acute/chronic bacterial infection require the systemic administration of antibiotics. Repeated administration of chemotherapeutic agents affects the testicular cells, accessory sex glands and may influence the andrological status of male animals. Various biochemical alterations in semen following repeated administration of some drugs have also been reported (Timmermans 1974). Enrofloxacin, a broad-spectrum fluoroquinolone group of antibiotic is routinely used for treatment of various bacterial infections. Alkaline phosphatase (AKP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are essential enzymes for metabolic process, which provide energy for survival, motility and fertility of spermatozoa. The concentration of AST and ALT in semen is a good indicator of semen quality because it measures sperm membrane stability (Pace and Graham 1970) and reflects the secretory activity of accessory sex glands. Thus increase in the percentage of dead spermatozoa in a semen sample causes high concentration of ALT and AST enzyme in the extracellular fluids due to sperm membrane damage and easy leakage of enzymes from spermatozoa. Furthermore, percentage of live spermatozoa in the semen is negatively correlated with AST level and AST/ALT ratio and semen quality (Daader et al. 1993; Taha et al. 2000). Alkaline phosphatase (AKP) and acid  phosphatase (ACP) were reported to have an important role on functional status of accessory sex glands, epididymis and ampullae (Ibrahim et al. 1985). In case of breeding bucks, the effect of antimicrobial therapy on these biochemical markers in semen is not yet studied. Keeping in view, the present study was undertaken with the objective to study the effect of enrofloxacin on biochemical parameters (AST, ALT, AKP and ACP) of seminal plasma of Barbari buck.

Materials and Methods

Six normal, healthy adult Barbari bucks (25-30 Kg) aged between 2 to 2.5 years stationed at the experimental sheds of Department of Veterinary Physiology, DUVASU, Mathura (U.P) were used as semen donors. The animals were grazed in a flock in Institute’s pasture daily from 9.00 AM to 3.00 P.M.., and 250g concentrate mixture having DCP 13% and TDN 69% was offered per animal daily with ad libitum watering. All the experimental animals were regularly dewormed for internal parasites.

The bucks were administered Enrofloxacin at the dose of 5mg/kg body weight intra-muscularly daily for 7 days. From each buck biweekly eighteen ejaculates (1st to 18th) were collected and one sample (0th) from each buck was collected before administration of Enrofloxacin using a non-oestrous doe as a dummy. The doner bucks were trained for semen collection prior to conduction of the experiment.

Seminal plasma was obtained after centrifuging semen at 3000 rpm for 15 minutes at 200C and activity of Aspartate aminotransferase (AST) and Alanine Aminotransferase (ALT) (Reitman and Frankel 1957), Acid Phosphatase (ACP) and Alkaline Phosphatase (AKP) (King 1965) were estimated, and total protein was estimated as suggested by Lowry et al (1951).           

Analysis of variance (ANOVA) was employed using computer software SPSS 13.0 (Statistical Package for Social Science) to compare enzyme activity before and after enrofloxacin administration in the seminal plasma of Barbari bucks.

Result and Discussion

The average activity of AST, ALT, AKP and ACP enzymes (Mean±SEM) in the neat seminal plasma of six Barbari bucks prior to and after drug administration ranged between 164.35 + 0.35 and 359.92 + 1.40 IU/L, 14.17 + 0.17 and 24.80 + 0.60 IU/L, 2409.98 + 0.95 and 3623.74 + 0.90 IU/L and 211.55 + 1.64 and 361.35 + 1.54 IU/L respectively. The total protein in the seminal plasma prior to and after drug administration ranged between 4.04 + 0.06 and 6.33 + 0.05 g/dl.

Following parentral administration of enrofloxacin the study on eighteen ejaculates (up to nine weeks) showed that there was a maximum and highly significant (P≤0.01) increase in AST level in seminal plasma of 9th ejaculate (Figure 1.2) whereas the maximum increase in ALT level was in seminal plasma of 5th ejaculate (Figure 1.1) as compared to ejaculate at 0 day. The AKP level increased significantly (P≤0.01) in the seminal plasma of 1st ejaculate and the increase was found to be maximum as compared to ejaculate at 0 day and it subsequently decreased up to 12th ejaculate and thereafter reached to its normal level (Figure 1.3). The ACP level in seminal plasma increased significantly (P≤0.01) to its maximum in 4th ejaculate as compared to ejaculate at 0 day which later decreased and reached its normal level in 18th ejaculate (Figure 1.4).

Figure 1.1 Effect of parentral administration of enrofloxacin on ALT level in seminal plasma

0 – Ejaculate prior to enrofloxacin administration

1 to 18 – Ejaculates after enrofloxacin administration

Figure 1.2 Effect of parentral administration of enrofloxacin on AST level in seminal plasma

0 – Ejaculate prior to enrofloxacin administration

1 to 18 – Ejaculates after enrofloxacin administration

Figure 1.3 Effect of parentral administration of enrofloxacin on AKP level in seminal plasma

0 – Ejaculate prior to enrofloxacin administration

1 to 18 – Ejaculates after enrofloxacin administration

 

The increase in ALT, AST, AKP and ACP levels in our study is in accordance with the findings of Verma (1996) who also found an increase in these enzymes in seminal plasma of buffalo bull administered with enrofloxacin. Higher levels of dead spermatozoa are associated with higher content of these enzymes in the seminal plasma (Kakar and Anand 1984; Kapila 1992). The increase in release of AST might be due to increased acrosomal abnormalities (Verma et al. 1999). The AST activity in the seminal plasma was associated positively with ALT, ACP, per cent dead spermatozoa but negatively with sperm concentration (Kale and Tomer 2000). Our results suggest  that increase in ALT, AST, AKP and ACP levels in seminal plasma may be due to increase in the number of dead and abnormal spermatozoa after administration of enrofloxacin. In our study the activity of ALT, AST, AKP and ACP in seminal plasma first increased and then decreased and reached its normal level, may be attributed to residual effect of enrofloxacin in time dependent manner. It may be

Figure 1.4 Effect of parentral administration of enrofloxacin on ACP level in seminal plasma

0 – Ejaculate prior to enrofloxacin administration

1 to 18 – Ejaculates after enrofloxacin administration

Figure 1.5 Effect of parentral administration of enrofloxacin on protein level in seminal plasma

0 – Ejaculate prior to enrofloxacin administration

1 to 18 – Ejaculates after enrofloxacin administration

 

concluded from this study that only after nine weeks of enrofolaxacin administration, the biochemical status of seminal plasma with respect to ALT, AST, AKP and ACP becomes normal.

References

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Ibrahim MAR and Boldizsar H. (1981). Studies on free amino acid content in seminal plasma of A. I. bulls of different performance. Acta. Vet. Academiae. Scienti. Arum. Hungaricae. 29: 263-269.

Kakar SS and Anand SR. (1984). Acrosomal damage and enzyme leakage during freeze preservation of buffalo spermatozoa. Indian Journal of Experimental Biology. 22: 5-10.

Kale MM and Tomer OS. (2000). Enzymatic constituents in crossbred buck semen. Indian Journal of Animal  Science. 70(1): 30-32

Kapila R. (1992). Leakage of enzymes during freezing of goat semen. M.Sc. Dissertation Submitted to NDRI, Karnal.

King J. (1965). Practical Clinical Enzymology. Pub. D. Van Nostrand Company (Canada) Ltd.25, Hollinger Road, Toronto16, p.104 & 301.

Lowry OH, Roserbroogh NS, Farr AL and Randal RJ. (1951). Protein measurement with folin phenol reagent. Journal of Biological Chemistry. 183: 265-275.

Pace MM and Graham EF. (1970). The release of Glutamic Oxaloacetic transaminase from Bovines spermatozoa as a test method for assessing semen quality infertility. Biology of Reproduction. 3: 140

Reitman S. and Frankel S. (1957). A colorimetric method for the determination of serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase. Americal Journal of Clinical Pathology. 28: 56-63

Taha TA, Abdel-Gawad EI and Ayoub MA. (2000). Monthly variation in some reproductive parameters of Barki and Awassi rams throughout 1year under sub-tropical conditions. Biochemical and enzymatic properties of seminal plasma. Animal  Science. 71: 325-332.

Timmermans L. (1974). Influence of antibiotics on spermatogenesis. Journal of Urology. 112: 348-349.

Verma HK. (1996). Studies on the effect of therapeutic agents on the seminal attributes of buffalo bulls. Ph.D. Dissertation Submitted to Punjab Agricultural University,Ludhiana.

Verma, HK, Pangawkar GR, Matharoo JS and Srivastava AK. (1999). Effect of enrofloxacin on the seminal attributes of buffalo bulls. Indian Veterinary Journal. 76: 1020-1022.

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