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Objective: To determine an optimum frequency of semen collection, so as to improve the efficacy of artificial insemination techniques and selective breeding programmes in duck farming. Methods:Thirty two ejaculates were collected by abdominal massage method from two groups of drakes, with 10 in each, at one day and two day intervals respectively. Routine seminal attributes such as volume, colour, pH, spermatozoa concentration, number of spermatozoa per ejaculate, individual motility, live sperm percentage, total sperm abnormalities and time taken to reduce Methylene blue (MBRT) were studied for comparison of their values in one day and two days of interval of semen collection. Twenty female laying ducks maintained for examining the fertility percent were regularly inseminated with 0.1 mL of neat pooled semen for fertility test. Results:The test of significance indicated a significant decrease (P<0.05) in the time taken to reduce methylene blue, a highly significant increase in the sperm concentration (P<0.01) and number of spermatozoa per ejaculation (P<0.01) and a non significant improvement in the seminal characteristics with respect to semen volume and mass motility was found in the ejaculates of two day interval than that of one day interval collections. Conclusion:Collection of semen at two days intervals is recommended for enhancing fertility in artificial insemination of duck. The methylene-blue reduction test, along with spermatozoa count and initial motility estimates were reported to be better indicator of semen quality.