Our objective was to determine the effects of a single treatment of human chorionic gonadotropin (hCG) or GnRH from d 5 to 7 of the estrous cycle on cycle length, expression of estrus and fertility in lactating dairy cows. Lactating Holstein cows (n = 354) located in Farm 1 and lactating Jersey cows located in Farm 2 (n = 210) detected in estrus by an Automated Activity Monitor (AAM) system from 27 to 50 days in milk (DIM) were randomly assigned to receive one of three treatments from d 5 to 7 of the estrous cycle: control (untreated; CON; Holstein, n = 111; Jersey, n = 66), GnRH (86 μg gonadorelin acetate im; Holstein, n = 116; Jersey, n = 75), or hCG (3,300 IU im; Holstein, n = 127; Jersey, n = 69). Ovaries were scanned with ultrasound in a random subgroup of cows (Holstein/Farm 1, n = 147; Jersey/Farm 2, n = 94) on the day of treatment and 3 or 4 d later to determine ovulation. Estrus was detected after treatment by an AAM, and peak activity and heat index were recorded. A random subgroup of cows observed in estrus after treatment received first AI from 51 to 80 DIM (Holstein, n = 208; Jersey, n = 138). Pregnancy diagnoses were performed by transrectal ultrasonography at 37 ± 3 d post-AI. Holstein and Jersey cows treated with GnRH and hCG had an increased (P < 0.05) ovulatory response compared with controls. Human chorionic gonadotropin decreased (74%; P = 0.05) and GnRH tended to reduce (75%; P = 0.07) the proportion of multiparous Holstein cows returning to estrus compared with CON (86%). Cows treated with hCG had a longer (P < 0.01) estrous cycle length (24.6 ± 0.3 d, Holstein; 23.0 ± 0.3 d, Jersey) compared with CON cows (22.7 ± 0.3 d, Holstein; 21.3 ± 0.3 d Jersey) and GnRH (22.9 ± 0.3 d, Holstein; 21.1 ± 0.3 d Jersey). The percentage of cows with high (≥80) peak activity and heat index did not differ (P > 0.50) between treatments, and milk production did not affect (P > 0.65) the duration of estrus. Pregnancy per AI (P/AI) was not affected by treatments in Holstein (P = 0.93; CON: 34.3%, GnRH: 35.4%, and hCG: 31.5%) and in multiparous Jersey cows (P = 0.35; CON: 34.3%, GnRH: 35.4%, and hCG: 31.5%), but hCG had greater (P = 0.03; 55%) P/AI than GnRH (30.0%) and a trend (P = 0.06) for greater P/AI than CON (33.3%) in primiparous Jersey cows. In summary, inducing the formation of an accessory corpus luteum from d 5 to 7 of the estrous cycle with hCG reduced expression of estrus in multiparous Holstein cows. Moreover, hCG increased estrous cycle length in Holstein and Jersey cows, and it did not affect first service P/AI at 37 ± 3 d post-AI in Holstein and multiparous Jersey lactating cows. However, hCG increased P/AI in primiparous Jersey cows. Future research with a larger number of cows is needed to confirm these intriguing fertility results.