Effects of oestrus synchronisation and fixed-time artificial insemination on the reproductive performance of dairy heifers

AIM: To compare the reproductive performance of heifers after oestrus synchronisation and fixed-time artificial insemination with non-synchronised heifers bred by herd sires. METHODS: Heifers from 10 spring-calving herds were randomly divided into two groups by herd, breed and age. Heifers in one group (the synchronised group, n = 478) were synchronised with a combination of progesterone, oestradiol benzoate and PGF2alpha, and inseminated 50-54 hours after progesterone treatment. Returns to first service were resynchronised with progesterone treatment 16-21 days after the fixed-time artificial insemination. Heifers in the other group (the control group, n = 470) did not receive any treatment and were bred by herd sires. RESULTS: The conception rate of synchronised heifers to the fixed-time artificial insemination was 51.2% and to the artificial insemination after resynchronisation 40.4%. The pregnancy rate at the end of the breeding season was lower (p<0.001) for the synchronised (92.9%) than for the control (97.2%) group. The interval from start of breeding to calving was earlier for synchronised (295.9 +/- 22.5 days, mean +/- s.d.) than for control (298.5 +/- 17.3 days) heifers. CONCLUSION: Results from this study indicate that the oestrus synchronisation programme used in the present study can reduce reproductive performance by increasing the empty rate compared with natural mating.     

Incorporation of estradiol benzoate to CIDR protocol improves the reproductive responses in crossbred dairy heifers

The present study was designed to determine the effect of estradiol benzoate (EB) on reproductive response following a controlled internal drug release (CIDR) protocol in crossbred (Sahiwal × Friesian) dairy heifers. In the first trial, a total of 100 crossbred dairy heifers were treated with CIDR protocol for 7 days and injected with the PGF_2α on day 6. After 24 h of CIDR removal, one group (EB?=?50) was injected with estradiol benzoate whereas the other (control?=?50) remained untreated. Estrus intensity and response were recorded visually and ovulation rate was recorded by ultrasonography. All heifers were artificially inseminated at 48 and 60 h following CIDR removal. Heifers were scanned for pregnancy within days 30–40 of artificial insemination (AI). In the second trial, two subgroups of heifers were included to observe the estrus and ovulatory events. The results of the first trial revealed that estrus response was achieved 100% in both the treatment groups. Estrus intensity (2.9?±?0.1 vs. 2.0?±?0.7) and ovulation rate (100 vs. 88%) differed significantly (P?<?0.05) between the EB and control groups. However, a tendency for higher pregnancy per AI was observed (54 vs. 36%; P?=?0.07) in EB than that in control groups. The results of the second trial revealed that a significantly (P?<?0.05) shorter estrus and earlier ovulatory events were observed in EB-treated heifers. It is concluded that the incorporation of estradiol benzoate to the CIDR protocol is helpful to improve the estrus signs and enhance the ovulation and the pregnancy per AI in crossbred dairy heifers.     

The effect of fixed-time artificial insemination protocol initiated at different stages of the estrous cycle on follicle development and ovulation in gilts

Hormonal products have been developed for fixed-time artificial insemination (FTAI) to improve the efficiency of swine production. Here, we evaluated the effect of an FTAI protocol initiated during different phases of the estrous cycle on follicle development and ovulation in gilts. A total of 36 gilts were equally divided into three groups designated as the luteal (L), follicular (F), and post-ovulation (O) groups and fed with 20 mg of altrenogest for 18 days, followed by intramuscular injection of 1000 IU PMSG at 42 h after withdrawal of altrenogest, and 100 μg of GnRH after an 80-h interval. The L group had the highest number of follicles 4–6 mm in diameter, as well as corpora hemorrhagica. The mRNA expression of caspase-9 in the L group were significantly lower than those in the O and F groups (P < 0.05), while CYP11A1 and VEGF mRNA expression levels were significantly higher (P < 0.05). Moreover, FSHR mRNA levels were significantly higher in the O group than in the L, F, and control groups (P < 0.05). LHCGR and CYP19A1 mRNA levels were the highest in the F group (P < 0.05). Thus, the changes in the expression of genes associated with follicular development, maturation, and ovulation identified in this study indicated that initiation of the FTAI protocol during the luteal phase induced a better environment for follicle development and ovulation in gilts.     

富铜酵母对西门塔尔牛发情周期生殖激素分泌的影响

选用16头18月龄、体况良好、体质量390 kg的西门塔尔牛育成母牛,采用完全随机分组设计分为4组,研究富铜酵母(0、8、16、24 mg/kg)对发情周期外周血清促黄体素、促卵泡素、孕酮和雌二醇分泌规律的影响.结果显示,8 mg/kg组和16 mg/kg组显著提高了发情周期促黄体素、促卵泡素、孕酮和雌二醇水平(P<0.05);24 mg/kg组有提高趋势,但与对照组比差异不显著(P>0.05).结果表明,添加富铜酵母以8～16 mg/kg对发情周期生殖激素分泌有促进作用,以8 mg/kg为佳,兼顾基础日粮的含铜量,建议日粮铜水平为13.87 mg/kg.     

Puncture and drainage of the subordinate follicles at timed artificial insemination prevents the risk of twin pregnancy in dairy cows

The objective of this study was to determine whether fluid drainage from the subordinate follicles by ovum pickup procedures prevents the risk of twin pregnancy without reducing the fertility of the cow. Lactating dairy cows with at least two follicles over 12 mm diameter located one on each ovary and selected from synchronized groups for fixed‐time insemination were assigned to a Control (n = 49) or Drainage (n = 49) group. The largest follicle was considered as the dominant follicle. Draining of all subordinate follicles (≥10 mm) was performed in the Drainage group. All drained follicles developed as a corpus luteum 7 days later. In the Control cows, the presence of two corpora lutea (55.1%) was lower (p = .003) than those in the Drainage cows (87.8%), whereas the incidence of twin pregnancies was 50% and 0% for the Control and Drainage groups, respectively (p < .001). Draining did not affect the pregnancy rate. These results indicate that puncture and drainage of the subordinate follicles at insemination may eliminate the risk of twin pregnancies and reduce the risk of subsequent pregnancy loss by increasing the incidence of additional corpora lutea.     

Synchronization of lactating dairy cows with prostaglandin F2 alpha: insemination on observed oestrus versus timed artificial insemination

The effect of two insemination policies after synchronization of oestrus on reproductive performance in two groups of cows was investigated. Oestrus was synchronized by two treatments with prostaglandin F2 alpha (PGF2 alpha) at a 14-day interval. Cows in Group 1 (n = 234) were inseminated twice on appointment 66 and 90 h after the second treatment. Cows in Group 2 (n = 222) were intensively watched for signs of oestrus after the synchronization protocol and inseminated on observed oestrus. Cows with abnormal discharge after synchronization were excluded from breeding and inseminated later during the study period. Service rate within 1 week after synchronization was higher in Group 1 than in Group 2 (89.3 versus 67.1%). Conception rates on first service did not differ between groups (33.2 versus 30.0%). Days to first service and days open were shorter in Group 1 (P < 0.05). The number of cows culled for infertility did not differ between the groups. Endometritis 14-20 days post-partum decreased the percentage of cows pregnant at the end of the study period in both groups but did not have a significant effect on conception rates and days open. It is concluded that additional inseminations required in the timed artificial insemination protocol were economically justified by the reduction in days open in comparison with insemination on observed oestrus after synchronization.     

LRH-A3 and HCG increase pregnancy rate during timed artificial insemination in dairy cows

The objective of this study was to use luteinizing hormone-releasing hormone A3 (LRH-A3) and human chorionic gonadotrophin (HCG) to improve pregnancy rate of dairy cows during timed artificial insemination (TAI). In experiment 1, the TAI process (0 d, GnRH, 100 μg; 7 d, PGF2α, 0.4 mg; 56 hr, GnRH, 100 μg; 16 hr, AI) was applied to 160 dairy cows on 50th and 60th days after parturition respectively. In experiment 2, 320 postpartum dairy cows were treated with TAI (Group A), TAI + 25 μg LRH-A3 (Group B), TAI + 1,500 IU HCG 5 days after AI (Group C), and TAI + 25 μg LRH-A3 + 1,500 IU HCG 5 days after AI (Group D). In experiment 3, endometrial cells were treated with HCG. The results showed that TAI did not affect the pregnancy rate, while LRH-A3 and HCG increased the pregnancy rate of the cow. HCG of 5 IU/ml and 10 IU/ml increased the expressions of leukemia inhibitory factor but decreased those of interleukin-6, epidermal growth factor and vascular endothelial growth factor in endometrial cells. This study provided a plan for the use of LRH-A3 and HCG to increase pregnancy rate during TAI in dairy cows.     

Effect of timing of artificial insemination after synchronization of ovulation on reproductive performance in Holstein dairy cows

The objective of this study was to determine the effect of timing of artificial insemination on pregnancy rates, calving rates, abortion rates, twinning rates, and calf gender ratio after synchronization of ovulation with Ovsynch protocol in Holstein dairy cows. The ovulation of 219 lactating Holstein dairy cows was synchronized using the Ovsynch protocol. Therefore, cows received an injection of GnRH followed by an injection of prostaglandin F_2α 7 days later and a second treatment with GnRH 2 days later. Cows were artificially inseminated at 0, 12, or 24 h after the second injection of GnRH. Reproductive performance did not differ between cows inseminated at 0 h (n?=?82), 12 h (n?=?66), or 24 h (n?=?71) after the last injection of GnRH (pregnancy rate: 0 h 48 %, 12 h 47 %, 24 h 52 %; abortion rate: 0 h 5 %, 12 h 0 %, 24 h 11 %; calving rate: 0 h 43 %, 12 h 47 %, 24 h 41 %; twinning rate: 0 h 2 %, 12 h 0 %, 24 h 0 %; calf gender ratio (F/M): 0 h 61:39 %, 12 h 48:52 %, 24 h 39:61 %; P?>?0.05). Pregnancy rates for cows inseminated in postpartum times of 50–75, 76–100, and >100 days within the first and ≥3 parities were statistically similar (P?>?0.05), but pregnancy rates for cows inseminated at different postpartum times of 50–75, 76–100, and >100 days within the second parity were different (P?<?0.01). In general, pregnancy rates of cows inseminated at different postpartum times (P?<?0.01) and parities (P?<?0.001) differed. The findings of the current study showed that rates of pregnancy, abortion, calving, and twinning of Holstein dairy cows subjected to artificial insemination at different times after synchronization were similar. These results also indicate that the timing of artificial insemination after synchronization did not influence calf gender ratio. Furthermore, pregnancy rates of Holstein dairy cows inseminated after synchronization were significantly influenced by postpartum time and parity number.     

Conception rates to fixed-time artificial insemination of two oestrus synchronisation programmes in dairy heifers

AIM: To evaluate the conception rate to fixed-time artificial insemination (FTAI) of two oestrus synchronisation programmes in dairy heifers on eight farms over 2 years.

METHODS: The study was conducted in 2008 and 2010 on eight farms near Palmerston North, New Zealand. Nulliparous Friesian and Friesian×Jersey heifers (13–15 months of age) were randomly allocated to one of two oestrus synchronisation programmes. Group 1 (GPG+P4; n=330), received gonadotrophin-releasing hormone (GnRH) I/M on Day 0, a progesterone (P4)-releasing intravaginal device from Days 0–7, prostaglandin F_2α (PGF) I/M on Day 7 and a second dose of GnRH at the time of FTAI on Day 9. The second group (P4+PGF; n=343) received a P4-releasing intravaginal device from Days 0–7, PGF on Day 6 and FTAI on Day 9. Pregnancy was diagnosed from Days 42–52 by transrectal ultrasonography.

RESULTS: The overall conception rate was 52.4% and 54.8% for the GPG+P4 and P4+PGF groups, respectively. The odds of conception for the two treatments were not different (OR=0.90; 95% CI=0.67–1.23), nor was there any difference between groups in different years (p=0.58). Farm affected conception rate (p=0.002), but there was no interaction with treatment (p=0.92) .

CONCLUSIONS: This study has shown that an alternative synchronisation programme can produce similar results in terms of conception rate to the GPG+P4 treatment, currently commonly used in heifers. More research is required to establish whether other modifications to the GPG+P4 programme can produce similar results at lower costs, and to identify and quantify farm factors which affect the economic benefit of heifer synchronisation.

CLINICAL RELEVANCE: This study indicated that synchronising heifers with P4 and PGF resulted in conception rates equivalent to those resulting from a GPG+P4 treatment, but with reduced drug costs. However, because heifers in the GPG+P4 group received the second GnRH injection at the time of AI, they needed only three yardings as opposed to the four required for the heifers treated with P4 and PGF. Thus, the choice of programme for an individual farm will depend on that farm's circumstances, in particular the cost of yarding the heifers.     

Comparison of two types of CIDR-based timed artificial insemination protocols for repeat breeder dairy cows

This study compared two types of controlled internal drug release (CIDR)-based timed artificial insemination (TAI) protocol for treatment of repeat breeder dairy cows. In the first trial of the experiment, 55 repeat breeder cows were randomly assigned to the following two treatments. (1) In the EB group, a CIDR device was inserted into the cows, and then the cows were administered an injection of 1 mg estradiol benzoate (EB) plus 50 mg progesterone (P4; Day 0). On Day 7, they were given an injection of PGF(2alpha) and the CIDR device was removed. The cows were given an injection of 1 mg EB on Day 8 and were subjected to TAI 30 h later (n=27). (2) In the gonadotrophin releasing hormone (GnRH) group, a CIDR device was inserted into the cows, and then the cows were administered an injection of 250 microg gonadorelin (GnRH; Day 0). On Day 7, they were given an injection of PGF(2alpha) and the CIDR device was removed. The cows were given an injection of 250 microg GnRH on Day 9 and were subjected to TAI 17 h later (n=28). In the second trial, 41 repeat breeder cows that were confirmed as not pregnant in the first trial were randomly assigned to the same two treatments used in the first trial (an EB group of 20 cows and a GnRH group of 21 cows). The ovaries of 15 cows from each group were examined by transrectal ultrasonography in order to observe the changes in ovarian structures, and blood samples were collected for analysis of serum P4 concentrations. The pregnancy rates following TAI in the first (18.5 vs. 32.1%) and second (40.0 vs. 38.1%) trials and the combined rates (27.7 vs. 34.7%) did not differ between the EB and GnRH groups. The proportions of cows with follicular wave emergence within 7 days did not differ between the EB (12/15) and GnRH groups (13/15). The interval to wave emergence was shorter (P<0.01) in the GnRH group than in the EB group, but there was no difference in the mean diameters of dominant follicles on Day 7 between the groups. Moreover, the proportions of cows with synchronized ovulation following a second EB or GnRH treatment did not differ between the groups. In conclusion, treatment with either EB or GnRH in a CIDR-based TAI protocol results in synchronous follicular wave emergence, follicular development, synchronous ovulation, and similar pregnancy rates for TAI in repeat breeder cows.     

Ovarian responses of dairy buffalo cows to timed artificial insemination protocol,using new or used progesterone devices,during the breeding season (autumn–winter)

This study evaluated the effect of new or used P4 devices on the ovarian responses of dairy buffalo that were administered an estradiol (E2) plus progesterone (P4)‐based timed artificial insemination (TAI) protocol during the breeding season. On the first day of the TAI protocol, 142 cows were randomly assigned to receive one of the following: a new device (New; 1.0 g of P4; n = 48); a device that had previously been used for 9 days (Used1x, n = 47); or a device that had previously been used for 18 days (Used2x, n = 47). Ultrasound was used to evaluate the following: the presence of a corpus luteum (CL); the diameter of the dominant follicle (ØDF) during protocol; ovulatory response; and pregnancies per AI (P/AI). Despite similar responses among the treatments, there was a significant positive association of the ØDF during TAI protocol with ovulatory responses and number of pregnancies. In conclusion, satisfactory ovarian responses and a satisfactory pregnancy rate were achieved when grazing dairy buffalo were subjected to the TAI protocol in breeding season, independent of whether a new or used P4 device was used. Furthermore, the presence of the larger follicle was associated with a higher ovulation rate and higher P/AI following TAI.     

The effects of progesterone treatment following artificial insemination on the reproductive performance of dairy cows

The objective of this study was to examine the effects of increasing circulating progesterone in multiparous Holstein cows, using an intravaginal progesterone implant between days 3 and 7 following fixed-time artificial insemination (FTAI). The animals were randomly assigned into two groups: (1) treatment group and (2) control (no treatment). The experiment took place during periods of high and mild temperatures. Pregnancy rates were evaluated by ultrasound at 30 and 60 days after FTAI; embryonic and fetal mortality rates were also evaluated on these dates. The temperature and humidity index on the day of FTAI were different (P?<?0.05) during the times of interest, but it did not affect conception rates. Neither the conception rates at days 30 and 60 nor the embryonic and fetal mortality rates differed between the groups during either the high- or mild-temperature periods (P?>?0.05). Throughout the experimental period, the serum progesterone concentration was higher in the treatment group than in the control (P?<?0.05). We conclude that the intravaginal progesterone implant inserted from days 3 to 7 following FTAI had no effect on conception rates at days 30 and 60 or on embryonic or fetal mortality rates but enhances the serum progesterone concentrations following treatment in multiparous Holstein cows during high- and mild-temperature periods.     

Efficacy of gonadotropin-releasing hormone administered at the time of artificial insemination of heifers and postpartum and repeat breeder dairy cows

The efficacy of gonadotropin releasing hormone (GnRH) on the fertility of dairy heifers and postpartum cows was studied. In one study, 300 cows were assigned randomly to 1 treatment from a 2 X 2 design, and 2 ml of 0.15M saline solution or 100 micrograms of GnRH was given IM on postpartum day (PPD) 14 and at the time of 1st postpartum breeding. In a 2nd study, repeat breeder cows (n = 346) were given saline solution or GnRH at the time of the 3rd breeding. In a 3rd study, heifers (n = 185) were given saline solution or GnRH at 1st breeding. Animals were observed for repeat estrus, and pregnancy diagnoses were made by rectal palpation 45 to 60 days after breeding. Conception rates were 15% to 18% higher for cows given GnRH, whether at PPD 14 or at the first postpartum breeding. When GnRH was given at the first breeding, conception rates were significantly higher (P less than 0.05). Conception rates for repeat breeder cows given GnRH were 25% higher than those in controls (P less than 0.005). Conception rates for heifers did not differ between treatments.     

同期发情定时输精技术对经产奶牛繁殖力的影响

为了研究同期发情定时输精技术对牛群体繁殖力的影响,试验选取同一牛场内不同单元的经产奶牛,随机分为2个组,分别采用自然发情输精和同期发情定时输精两种方法对奶牛进行人工输精,比较两种方法对奶牛群体繁殖力的影响。研究结果显示:同期发情组和自然发情组的参配率分别为,产后54 d之前为96.3%VS 12.1%,55～90 d为64.6%VS 44.6%,91～130 d为48.1%VS 50.1%,131～170 d为39.9%VS 38.7%,180 d内的总参配率分别为249.1%VS 145.5%;相应时段的同期发情组的受胎率为39.2%、36.7%、36.5%、33.9%、92.6%,自然发情组的受胎率为44.8%、47.3%、46.2%、46.1%、 67.5%;2个组的年繁殖率分别为93.8%VS 84.6%、空怀天数为(91.6±18.3)d VS (131.7±26.8)d、配种次数分别为2.5 VS 1.8、淘汰率分别为29.1%VS 13.4%。以上结果表明,在奶牛生产中,使用同期发情定时输精技术,能显著提高牛群的繁殖力,缩短空怀时间。     

Pregnancy rate after timed artificial insemination in early post-partum dairy cows after Ovsynch or specific synchronization protocols

The present study was designed to compare the reproductive performance of pre-synchronized post-partum dairy cows subjected, either to the Ovsynch protocol without screening for ovarian status (control group), or to a specific oestrous synchronization protocol applied according to their ovarian status, as determined by transrectal ultrasound (experimental group). The study was conducted on 428 lactating dairy cows. Cows in the Ovsynch group (n = 205) were synchronized and time inseminated after receiving the Ovsynch protocol treatment. Cows in the specific synchronization (Ssynch) group (n = 223) were weekly subjected to transrectal ultrasound exams for 4 weeks, or until AI or starting treatment, and divided into four subgroups according to their ovarian status: (i). corpus luteum (CL) subgroup (n = 130), cows with a CL; (ii). natural oestrus (NE) subgroup (n = 58), cows showing NE; (iii). anovulatory follicles (AF) subgroup (n = 26), cows considered to have AF; and (iv). ovarian cysts (OC) subgroup (n = 9), cows with OC. Cows in the Ssynch group were synchronized and time inseminated following a specific oestrous synchronization protocol, or inseminated at NE. Logistic regression analysis was carried out for the dependent variables ovulation and pregnancy rates to first and to second AI (second AI: first AI + return AI). Cows subjected to Ssynch were 2.1 times more likely to become pregnant at first and at second AI compared with those synchronized using the Ovsynch protocol (P < 0.0001). Our results show that the response of post-partum pre-synchronized cows to a specific oestrous synchronization protocol applied according to their ovarian status is more effective than the response to the Ovsynch protocol applied without taking into account the ovarian status of the animals.     

Relationship of the conception rate and the side (left or right) of preovulatory follicle location at artificial insemination in dairy heifers

In this study, we examined the locational effect (left or right ovary) of the preovulatory follicle (PF) on fertility in dairy heifers. In total, 1,111 artificial inseminations (AI) were analyzed. At AI, PF locations were examined using rectal palpation, and heifers were divided into two groups on their PF locations: (i) the PF located in the left ovary (L‐PF); and (ii) the PF located in the right ovary (R‐PF). Pregnancy was diagnosed by rectal palpation 60 days after AI. The conception rate was 50.7% in all heifers. Conception rate was significantly higher in the L‐PF (60.1%) than in the R‐PF (46.2%). The conception rate was significantly lower by sexed semen (48.6%) than conventional semen (59.1%). Conception rates divided by the semen type (sexed: n = 896, conventional: n = 215) were significantly higher in the L‐PF than in the R‐PF for both semen types (sexed; L‐PF vs. R‐PF: 57.3% vs. 44.4%, conventional; L‐PF vs. R‐PF: 72.3% vs. 53.3%). In addition, season, age, AI number, and the number of re‐inseminations at the same estrus did not affect conception rates. In summary, PF development in the left ovary was associated with increased conception rates in dairy heifers.     

Fertility of Angus cross beef heifers after GnRH treatment on day 23 and timing of insemination in 14‐day CIDR protocol

This study compared artificial insemination pregnancy rate (AI‐PR) between 14‐day CIDR‐GnRH‐PGF2α‐GnRH and CIDR‐PGF2α‐GnRH synchronization protocol with two fixed AI times (56 or 72 hr after PGF2α). On day 0, heifers (n = 1311) from nine locations assigned body condition score (BCS: 1, emaciated; 9, obese), reproductive tract score (RTS: 1, immature, acyclic; 5, mature, cyclic) and temperament score (0, calm; and 1, excited) and fitted with a controlled internal drug release (CIDR, 1.38 g of progesterone) insert for 14 days. Within herd, heifers were randomly assigned either to no‐GnRH group (n = 635) or to GnRH group (n = 676), and heifers in GnRH group received 100 μg of GnRH (gonadorelin hydrochloride, IM) on day 23. All heifers received 25 mg of PGF2α (dinoprost, IM) on day 30 and oestrous detection aids at the same time. Heifers were observed for oestrus thrice daily until AI. Within GnRH groups, heifers were randomly assigned to either AI‐56 or AI‐72 groups. Heifers in AI‐56 group (n = 667) were inseminated at 56 hr (day 32 PM), and heifers in AI‐72 group (n = 644) were inseminated at 72 hr (day 33 AM) after PGF2α administration. All heifers were given 100 μg of GnRH concurrently at the time AI. Controlling for BCS (p < .05), RTS (p < .05), oestrous expression (p < .001), temperament (p < .001) and GnRH treatment by time of insemination (p < .001), the AI‐PR differed between GnRH treatment [GnRH (Yes – 60.9% (412/676) vs. No – 55.1% (350/635); p < .05)] and insemination time [AI‐56 – 54.6% (364/667) vs. AI‐72 – 61.8% (398/644); (p < .01)] groups. The GnRH treatment by AI time interaction influenced AI‐PR (GnRH56 – 61.0% (208/341); GnRH72 – 60.9% (204/335); No‐GnRH56 – 47.9% (156/326); No‐GnRH72 – 62.8% (194/309); p < .001). In conclusion, 14‐day CIDR synchronization protocol for FTAI required inclusion of GnRH on day 23 if inseminations were to be performed at 56 hr after PGF2α in order to achieve greater AI‐PR.     

Cytological endometritis diagnosed at artificial insemination in repeat breeder dairy cows

This study aimed to (i) assess the prevalence of cytological endometritis (CYTO) diagnosed at artificial insemination (AI); (ii) evaluate the effect of CYTO on the pregnancy outcome of the same AI sample; and (iii) determine the risk factors associated with CYTO diagnosed at AI in repeat breeder (RB) dairy cows. We analysed the productive and reproductive performances of 146 RB Holstein‐Friesian cows. To obtain a CYTO sample at AI, we used the cytotape technique. Generalized mixed effect models were computed to find the risk factors associated with the pregnancy and CYTO outcome. Based on ≥1% PMN cut‐off point, the CYTO prevalence at AI in RB cows was 25.3%. The overall pregnancy at AI was 44.2%. The conception rate in CYTO‐positive (n = 37) RB cows was 29.7% versus 49.5% for CYTO‐negative (n = 109) cows. A RB cow diagnosed CYTO positive at AI had 0.47 [odds ratio (OR)] odds to become pregnant in comparison with a CYTO‐negative cow. Cows that produced more milk than their counterparts in this study had increased odds (OR = 1.01) to be CYTO positive at AI. A novel risk factor positively associated with CYTO diagnosed at AI in RB cows was the level of daily milk urea (OR = 1.11). To conclude, CYTO at the moment of AI had a significantly negative effect on the pregnancy outcome in RB dairy cows. However, as only one of fourth of RB cows is affected with CYTO at AI, it may not be considered a key element associated with the RB syndrome.