This paper deals with the study on comparative efficacy and in vitro activity of cow urine and cow dung for controlling root rot disease of cucumber caused by Fusarium solani f. sp. cucurbitae Snyder & Hansen following slide germination and mycelial growth inhibition tests. Results showed that both germination of conidia and the percentage inhibition of mycelial growth decreased or suppressed and varied greatly with respect to different hour and days of incubation and kind of bio-matters. In between two bio-matters cow urine was found more effective than that of cow dung in conidial germination. No germination of conidia was recorded after one hour of incubation in any medium whereas in cow urine germination of conidia was not also observed even after 2 hours of incubation. After 7 hours of incubation out of 200 conidia of F. solani f. sp. cucurbitae, 28 in cow urine and 64 in cow dung were germinated while in control a total germinated conidia was 185. In case of percentage inhibition of conidial germination the highest percentage(100%) was recorded in cow urine after 2 hours of incubation followed by 3 hours(96.0%), 4 hours(91.0%) and 6 hours(89.4%). During the test on inhibition of mycelial growth, the highest percentage(62.8%) was recorded in cow urine potato dextrose agar(CUPDA) medium tested after 4 days of incubation, followed by 3 days(60.5%), 5 days(56.5%) and 2 days(55.0%). In this test cow dung potato dextrose agar(CDPDA) had less efficacy in suppression of the percentage inhibition of mycelial growth.