The occurrence of ovarian follicular cyst has significant effects on the reproductivity in cattle and lead to economic loss. The objective of this study is to investigate the efficacy of hormonal therapy for treating follicular cysts to improve the fertility in crossbred cows and the application of ultrasound guided transvaginal follicular aspiration technique in non-responsive chronic follicular cysts. A total of 28 crossbred cows with follicular cysts were included in the study and were treated with GnRH analogue @ 20μg intramuscularly on day 0. The cows which responded to first GnRH injection (16/28, 57.14%) having corpus luteum on day 11 were termed as Group I (n=16). Group I cows were treated with Co-synch protocol. The cows which did not respond to first GnRH having persistent cyst on day 11 were termed as group II (n=12) and received second GnRH injection. Only seven cows responded for second GnRH injection (7/12, 58.33%) and the rest of the protocol was followed similar to that of group I. Three among the five cows exhibited estrus and were re-inseminated. Ultrasound guided transvaginal follicular aspiration was performed for remaining five non-responsive cows in group II. Pregnancy diagnosis on day 60 post AI revealed nine out of sixteen cows were pregnant (9/16, 56.25%) in group I and three out of seven were pregnant (3/7, 42.85%) in group II responded cows. However, the recurrence of fluid accumulation in the follicular cyst were recorded in two cows (2/5, 40.00%) and three cows had recovered (3/5, 60.00%) following follicular aspiration and later one cow was pregnant. The overall pregnancy rate in group I, group II, hormonal therapy and follicular ablation were 56.25%, 33.33%, 52.17% and 20.00%, respectively. The ovarian follicular cysts were characterized by higher levels of BUN, calcium and lower levels of glucose, cholesterol, total protein, albumin, creatinine, AST and phosphorus. Hence, in the cases of failure of hormonal therapy, ultrasound guided transvaginal follicular aspiration technique can be advocated for the management of chronic and non-responsive cystic ovarian follicles in dairy cattle.