Factors associated with estrous expression and subsequent fertility in lactating dairy cows using automated activity monitoring

The objective of this observational study was to identify factors associated with estrous duration (DU) and intensity measured as the peak of activity (PA) change and subsequent fertility in lactating Holstein cows using a neck-mounted automated activity monitor (Heatime Pro, SCR Engineers Ltd., Netanya, Israel). Ambient temperature and relative humidity were recorded hourly to calculate the temperature-humidity index (THI). A total of 5,933 estrus events from 3,132 cows located on 8 commercial dairy farms in Germany were used for this study. Farms participated in monthly DHIA testing. Pregnancy diagnosis was performed either by transrectal palpation [farm 1: 42 ± 3 d; farm 3: 40 ± 3 d; farms 4 and 8: 38 ± 3 d; farm 5: 43 ± 3 d after artificial insemination (AI)] or transrectal ultrasonography (farms 2, 6, and 7: 30 ± 3 d after AI). Estrous intensity was categorized based on peak activity of estrus into low (35–89 index value), and high (90–100 index value) PA. Overall, 73.5% of estrus events were of high PA. The mean (± standard error of the mean) DU was 14.94 ± 0.06 h. There was a strong correlation between DU and PA (r = 0.67). In the final statistical model, only PA was associated with pregnancy per artificial insemination (P/AI), with 1.35 greater odds of pregnancy for cows with high PA compared with cows with low PA. Increased THI 1 wk before AI was associated with shorter DU, lower PA, and decreased P/AI. A small percentage of cows (4.7%) showed short interestrus intervals (i.e., more than 1 activity peak within 7 d close to the event of estrus), resulting in reduced DU, PA, and P/AI. The change of weighted rumination was associated with DU and PA, as a lower nadir was associated with a greater risk for high PA and long DU. There was no association, however, between the nadir of change of weighted rumination and P/AI. Whereas milk yield and somatic cell count from the DHIA test date before AI were negatively associated with estrous expression, neither milk yield nor somatic cell count was associated with P/AI. Surprisingly, multiparous cows expressed estrus with longer DU (13.15 ± 0.31 h) compared with primiparous cows (12.52 ± 0.32 h), whereas PA did not differ among parities. Pregnancy per AI was greater for primiparous (29.4%) than for multiparous (22.1%) cows. An estrus event with long DU or high PA was more likely later in lactation. Milk fat, milk protein, milk urea nitrogen, and lactose from the DHIA test date closest to AI had no association with estrous expression or P/AI. In conclusion, DU and PA were highly correlated, and cows with high PA were particularly associated with greater odds for pregnancy. A negative association between estrous expression and P/AI was identified for increased THI 1 wk before AI and cows with short interestrus intervals using automated activity monitor.     

Comparison of ovarian function and circulating steroids in estrous cycles of Holstein heifers and lactating cows

Ovarian function was compared between nulliparous heifers (n = 29; 10 to 16 mo old) and lactating Holstein cows (n = 31; 55.9 +/- 3.5 d postpartum). Follicular dynamics, corpus luteum growth, and regression, and serum steroid concentrations were evaluated through ultrasonography and daily blood sampling. Most heifers (27 of 29) but only 14 of 31 cows had typical spontaneous estrous cycles after cycles were initiated. Twelve cows had atypical cycles, and 5 became anovulatory during the study. The 12 cows with atypical estrous cycles had low serum estradiol after luteolysis and failed to ovulate the dominant follicle present at luteolysis. Heifers and cows with typical cycles were compared directly. Interovulatory intervals were similar between heifers (22.0 +/- 0.4 d) and cows (22.9 +/- 0.7 d). Those animals had estrous cycles with either 2 (15 heifers; 11 cows), 3 (9 heifers; 2 cows), or 4 follicular waves (3 heifers; 1 cow). Cows ovulated later after luteolysis than heifers (5.2 +/- 0.2 vs. 4.6 +/- 0.1 d, respectively), and had more multiple ovulations (17.9 vs. 1.9%). Maximal serum estradiol concentration preceding ovulation was lower in cows than in heifers (7.9 +/- 0.8 vs. 11.3 +/- 0.6 pg/mL) even though ovulatory follicles were larger in cows (16.8 +/- 0.5 vs. 14.9 +/- 0.2 mm). Similarly, maximal serum progesterone concentration was lower for cows (5.6 +/- 0.5 vs. 7.3 +/- 0.4 ng/mL), whereas maximal volume of luteal tissue was larger for cows than heifers (11,120 +/- 678 vs. 7303 +/- 308 mm3). Thus, higher incidence of reproductive anomalies in lactating cows, such as low conception rate, ovulation failure, delayed ovulation, and multiple ovulations, may be due to lower circulating steroid concentrations in spite of larger ovulatory follicles and luteal structures.     

Association of estrous expression detected by an automated activity monitoring system within 40 days in milk and reproductive performance of lactating Holstein cows

The objective of this observational study was to evaluate the association of estrous expression within 40 days in milk (DIM) using a neck-mounted automated activity monitor (Heatime Pro; SCR Engineers Ltd.) with reproductive performance in lactating Holstein cows. A total of 2,077 cows (614 primiparous cows and 1,463 multiparous cows) from 5 commercial dairy farms were included in the statistical analyses. Activity data from the first 7 d after calving were excluded. An estrus event was defined as an activity change index ≥35 for more than 2 h. Cows were classified according to the number of estrus events from d 7 until d 40 postpartum into 3 categories: (1) no estrus event (Estrus0); (2) one estrus event (Estrus1), and (3) 2 or more estrus events (Estrus2). Generalized linear mixed models were used to analyze continuous and categorical data. Shared frailty models were used for time to event data. Overall, 52.7% of cows had no estrus event detected by an automated activity monitor system from d 7 until d 40 postpartum. Herd level prevalence of Estrus0 ranged from 37.5 to 58.4%. Estrous expression from d 7 until d 40 postpartum affected estrous duration and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest duration (13.2 ± 0.33 h) compared with cows in Estrus1 (13.8 ± 0.36 h) and Estrus2 (14.8 ± 0.41 h). Cows in Estrus2 had a longer estrous duration at first postpartum AI compared with cows in Estrus1. Among Estrus0 cows, 46.2% had an estrus event with high intensity at first postpartum AI. Among cows in Estrus1 and Estrus2, 50.8 and 53.8% had an estrus event with high intensity at first postpartum AI, respectively. There was a significant difference between Estrus2 and Estrus0 and a tendency between Estrus0 and Estrus1. There was no difference between Estrus1 and Estrus2. For Estrus0, Estrus1, and Estrus2 cows, pregnancy per AI was 29.4, 30.9, and 37.8%, respectively. There was a significant difference between Estrus0 and Estrus2 and Estrus1 and Estrus2. There was no difference between Estrus0 and Estrus1. Estrous expression from d 7 until d 40 postpartum affected time to first AI and time to pregnancy. Compared with Estrus0 cows, cows in Estrus1 [hazard risk (HR) = 1.74] and Estrus2 (HR = 1.77) had an increased hazard of being inseminated within 100 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to first AI were 70, 59, and 58 for cows in Estrus0, Estrus1, and Estrus2, respectively. Compared with Estrus0 cows, cows in Estrus1 (HR = 1.28) and Estrus2 (HR = 1.33) had an increased hazard of becoming pregnant within 200 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to pregnancy were 127, 112, and 103 for Estrus0 cows, Estrus1 and Estrus2, respectively. In conclusion, cows with no estrous expression from 7 to 40 DIM had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity.     

The relationship between estrous behavioral score and time of ovulation in dairy cattle

One of the major contributors to poor fertility of a dairy herd is ineffective detection of estrus. Recently, it has become evident that cow factors contribute largely to low detection rates. Until now, standing behavior has been the symptom used to determine the right moment for insemination. However, standing behavior is not observed in over 50% of the cows in estrus in a number of herds, and no data are available to relate the moment of ovulation to other estrous symptoms. Therefore, in the present study, cows were observed for a number of visual signs of estrus, twice per day for 30 min, and the moment of ovulation was estimated by transrectal ultrasonographic scanning of the ovaries. In this study, 100 cows were detected in estrus visually, using a scoring system. Standing behavior was observed in 50% of those cows, although there was more than one cow in estrus at a time in most (85%) cases. There was no correlation between follicular size and ovulation time or estrous behavior score. Levels of 305-d milk yield and parity were also not correlated with estrous behavior scores. However, a significant correlation (0.31) was found between detection score and day of ovulation. A retrospective analysis revealed that cows that ovulated 0 to 24 h after artificial insemination (AD) scored almost three times the number of estrous behavior points compared to the ones that ovulated 24 to 48 h after AI. Ovulation more than 48 h after AI resulted in pregnancy in only 15% of the cows.     

Production,metabolism, and follicular dynamics in multiparous dairy cows fed diets providing different amounts of metabolizable protein prepartum and postpartum

Our objectives were (1) to determine whether increasing metabolizable protein (MP) supply above requirements in late-gestation cows would benefit health, milk production, and reproduction; (2) to determine whether an increased supply of MP postpartum affects production; and (3) to determine whether supply of MP prepartum interacts with MP supply postpartum. Pregnant nonlactating cows (n = 60) blocked by expected parturition date were assigned to 1 of 3 prepartum diets from 21 d prepartum to parturition: 12% crude protein (CP) soybean meal (SBM) supplement (LSB); 15% CP SBM supplement (HSB); and 15% CP SBM plus animal-marine protein supplement (HMP). Diets were formulated to supply an estimated 924, 988, and 1,111 g/d of MP, respectively, at 11.5 kg of dry matter intake (DMI). After parturition, cows received diets containing 18% CP, either from SBM (SB) or SBM plus animal-marine protein (AMP) supplements, that provided 2,056 (SB) or 2,293 g/d (AMP) of MP at 21 kg of DMI; thus, treatments were in a 3 × 2 factorial arrangement. Milk production and DMI were recorded for 63 d postpartum. Prepartum DMI was lower at wk ?3 for cows fed LSB compared with those fed HSB or HMP. Postpartum DMI did not differ significantly between cows fed SB and those fed AMP (20.8 vs. 19.6 kg/d). Milk production did not differ due to prepartum diets or postpartum diets. Milk fat and protein percentages were not affected by prepartum or postpartum diets. Cows fed AMP postpartum tended to produce more milk fat, but 4% fat-corrected milk (FCM) did not differ from SB-supplemented cows (33.6 kg/d vs. 32.2 kg/d). Gross feed efficiency (FCM/DMI) was greater for cows fed AMP postpartum (1.82 vs. 1.68). Prepartum concentrations of urea N in plasma were lower for LSB than for HSB and HMP, and HSB was greater than HMP. Postpartum concentrations of nonesterified fatty acids and β-hydroxybutyrate were greater for cows fed AMP postpartum than for those fed SB. Postpartum urea N was higher for SB than for AMP (14.4 vs. 12.5 mg/dL). Concentration of total protein in plasma was greater postpartum for cows fed HSB or HMP prepartum than for those fed LSB, and was greater postpartum for cows fed AMP than for those fed SB. Hepatic concentrations of total lipids and triglyceride did not differ among treatments. Hepatic glycogen was greater postpartum for cows fed SB postpartum. Feeding HSB or HMP increased the number of follicles 6 to 9 mm in diameter compared with LSB. The size of the largest follicle was increased by HMP compared with HSB. In conclusion, increasing the amount of MP fed to cows during the last 21 d prepartum did not affect milk production or BCS but increased plasma total protein concentration. Follicular dynamics were improved by increasing the amount of MP prepartum. Feeding HMP prepartum improved follicular dynamics prepartum and increased milk fat yield in wk 1. Feeding AMP postpartum increased efficiency of FCM production and plasma total protein. We found few interactions between prepartum and postpartum MP supply.     

Anogenital distance is associated with postpartum estrous activity,intensity of estrous expression,ovulation, and progesterone concentrations in lactating Holstein cows

The objectives of this retrospective observational study were to determine the associations of anogenital distance (AGD) with (a) postpartum estrous activity, (b) diameter of the preovulatory follicle, (c) intensity of estrous expression, (d) postestrus ovulation, (e) corpus luteum (CL) size, and (f) concentrations of progesterone at estrus and on d 7 after estrus. Lactating Holstein cows (n = 178; 55 primiparous, 123 multiparous) were enrolled into the study during the first postpartum week. All cows were continuously monitored by a pedometer-based automated activity monitoring (AAM) system for estrus. Postpartum estrous activity was assessed using the AAM estrus alerts, in which cows with at least one true estrus alert (i.e., a relative increase in steps from each cow's baseline detected by the AAM and the presence of at least one follicle >15 mm, a CL <20 mm, or no CL detected by ultrasound) by the first 50 d in milk (DIM) were considered to have commenced estrous activity. At the estrus alert >60 DIM, ovulation was determined by ultrasound at 24 h, 48 h, and 7 d after estrus, and blood samples were collected at estrus alert and on d 7 after estrus for progesterone analysis. The AGD was measured from the center of the anus to the base of the clitoris and classified as either short- or long-AGD using 2 cut-points of 148 mm (predictive of the probability of pregnancy to first insemination; short-AGD, n = 115; long-AGD, n = 63) and 142 mm (the median AGD; short-AGD, n = 90; long-AGD, n = 88). Regardless of the cut-point used, early postpartum estrous activity by 50 DIM (67 vs. 54%), duration of estrus (11.6 vs. 9.7 h), and preovulatory follicle diameter (20 vs. 19 mm) were greater in short-AGD than in long-AGD cows. Increased peak of activity at estrus in short-AGD cows (354 vs. 258% mean relative increase) was affected by an interaction between AGD and parity in which multiparous long-AGD cows had lesser relative increase in activity than primiparous cows (217 vs. 386%, respectively). Mean progesterone concentration at estrus was lesser in short-AGD (0.47 vs. 0.61 ng/mL) than in long-AGD cows. The ovulatory response at 24 h did not differ, but at 48 h (91 vs. 78%) and on d 7 after estrus (97 vs. 84%) it was greater in short-AGD cows. Although CL diameter on d 7 after estrus did not differ, short-AGD cows had greater progesterone concentration 7 d after estrus than long-AGD cows (4.1 vs. 3.2 ng/mL, respectively). In conclusion, greater proportions of short-AGD cows commenced estrous activity by 50 DIM, had larger preovulatory follicles, exhibited greater duration of estrus, had reduced progesterone concentration at estrus, had greater ovulation rates and progesterone concentration 7 d after estrus compared with long-AGD cows, with no difference in CL size between AGD groups. Because all the differences in physiological characteristics of short-AGD cows reported herein favor improved reproductive outcomes, we infer that these are factors contributing to improved fertility reported in short-AGD cows compared with long-AGD cows.     

Occurrence and greater intensity of estrus in recipient lactating dairy cows improve pregnancy per embryo transfer

The aim of this study was to determine the association between occurrence and intensity of estrous expression with pregnancy success in recipient lactating dairy cows subjected to embryo transfer (ET). Two observational studies were conducted. Holstein cows were synchronized using the same timed ET protocol, based on estradiol and progesterone in both experiments. At 9 d after the end of the timed ET protocol only animals that had ovulated were implanted with a 7-d embryo [experiment 1 (Exp. 1); n = 1,401 ET events from 1,045 cows, and experiment 2 (Exp. 2); n = 1,147 ET events from 657 cows]. Embryos were produced in vivo (Exp. 1 and Exp. 2) and in vitro (only Exp. 2), then transferred to recipient cows as fresh or frozen-thawed. Pregnancy was confirmed at 29 and 58 d after the end of timed ET protocol. In Exp. 1, animals had their estrous expression monitored through a tail chalk applied on the tail head of the cows and evaluated daily for chalk removal (no estrus: 100% of chalk remaining; estrus: <50% of chalk remaining). In Exp. 2, cows were continuously monitored by a leg-mounted automated activity monitor. Estrous expression was quantified using the relative increase in physical activity at estrus in relation to the days before estrus. Estrous expression was classified as no estrus [<100% relative increase in activity (RI)], weak intensity (100–299% RI), and strong intensity (≥300% RI). Data were analyzed by analysis of variance using mixed linear regression models (GLIMMIX) in SAS (SAS Institute Inc.). A total of 65.2% (914/1,401) and 89.2% (1,019/1,142) of cows from Exp. 1 and Exp. 2, respectively, displayed estrus at the end of the ovulation synchronization protocol. In Exp. 1, cows expressing estrus before to ET had greater pregnancy per ET than those that did not [41.0 ± 2.3% (381/914) vs. 31.5 ± 2.9% (151/487), respectively]. Similarly, in Exp. 2, cows classified in the strong intensity group had greater pregnancy per ET compared with cows in the weak intensity and no estrus groups [41.3 ± 2.2% (213/571) vs. 32.7 ± 2.7% (115/353) vs. 11.3 ± 3.5% (26/218), respectively]. There was no effect of ET type on pregnancy per ET in Exp. 1. However, in Exp. 2, cows that received an in vivo-produced embryo, either fresh or frozen, had greater pregnancy per ET compared with cows that received in vitro-produced embryo. Cows receiving embryos in the early blastocyst and blastocyst stage had greater fertility compared with cows receiving embryos in the morula stage. There was an interaction between the occurrence of estrus and the stage of embryo development on pregnancy per ET, cows which displayed estrus and received a morula or early blastocyst had greater pregnancy per ET than cows that did not display estrus. In conclusion, the occurrence and the intensity of estrous expression improved pregnancy per ET in recipient lactating dairy cows and thus could be used as a tool to assist in the decision making of reproduction strategies in dairy farms.     

Ovarian follicular activity in lactating Holstein cows supplemented with monensin

The objective of this study was to determine effects of monensin on ovarian follicular development and reproductive performance in postpartum dairy cows. Forty-eight multiparous Holstein cows were randomly assigned to receive either a control total mixed ration (n = 24) or the same diet plus 22 mg of monensin/kg (n = 24) from 21 d before anticipated calving until cows were either confirmed pregnant or were >180 d postpartum. Monensin had no effect on development of the first dominant follicle postpartum or the numbers of class 1 (3 to 5 mm), 2 (6 to 9 mm), or 3 (10 to 15 mm) follicles. Control cows had more class 4 (>15 mm) follicles at 10 to 13 d postpartum than cows in the monensin group. The first dominant follicle postpartum ovulated, regressed, or became cystic unrelated to differences between diets. However, the first ovulation postpartum occurred earlier in monensin-fed cows than in the control group (27.2 +/- 2.1 d vs. 32.4 +/- 1.5 d), with no dietary effects on the diameter of the ovulating follicle. Similarly, treatments did not differ in the proportion of cows with 2 or 3 waves of ovarian follicular development per cycle, nor in the number of follicles of all classes during the breeding period. Times of ovulation following treatment with prostaglandin F2alpha were not different between dietary groups. Pregnancy rates after timed artificial insemination were similar between diets. Supplementation with monensin resulted in a shorter postpartum interval to first ovulation but did not affect other reproductive measures in healthy, lactating dairy cows.     

Effect of dietary protein content on estrous behavior of dairy cows during early and mid lactation

One of the main contributing factors to the decline in fertility in contemporary dairy farming is the inability to detect cows in estrus. In the current study, 90 Holstein dairy cows [45 primiparous and 45 multiparous (mean parity of 3.1)] were allocated to 1 of 3 treatments at calving; 173, 144, or 114 g of crude protein/kg of dry matter. Estrous behavior was recorded for one 30-min period every 12 h from calving until all animals reached 140 d postpartum. Behavioral activities were recorded according to a scoring system developed by Van Eerdenburg et al. (1996), with 9 key estrous behavioral activities each allocated a given number of points. If the total score allocated was greater than or equal to 50 points during a single or consecutive observational periods, then the animal was deemed to be in estrus. A total of 238 estrous cycles scored 50 points or above on the Van Eerdenburg et al. (1996) scale in this experiment, with 51.7% of these cycles being characterized as standing immobile on mounting. There were no direct effects of dietary protein content on estrous behavior; however, 3 significant stage of lactation × protein treatment interactions occurred for the behavioral activities (mucous discharge, chin resting, and mounting the head side of another cow), but no consistent trends were apparent from the predicted means. There was a significant influence of parity on the frequency of mounting the head side of another cow and total number of behavior activities displayed per estrous cycle. In both cases multiparous animals displayed fewer behavioral activities than primiparous animals. An increase in the size of the sexually active group (animals in estrus at the same time, up to 5) significantly increased the expression of mounting or attempting to mount another cow, the number of cycles in which standing immobile on being mounted was observed, the total estrous score and the proportion of cyclic animals that were diagnosed as being in estrus. The most frequent behavioral activity displayed was chin resting (89.5% of cycles), and the most reliable behavior was standing immobile on mounting (when expressed 96.4% of cows were in estrus). The most dependable (function of reliability and frequency displayed) sign of estrus was mounting or attempting to mount another cow. This behavior was expressed in 83% of cycles, and when expressed, 89% of animals were in estrus.     

Effect of transition cow health and estrous expression detected by an automated activity monitoring system within 60 days in milk on reproductive performance of lactating Holstein cows

The objective of this observational study was to evaluate the association of transition cow health and estrous expression, detected by an automated activity monitoring system (Smarttag Neck, Nedap Livestock Management), with reproductive performance in lactating Holstein cows. A total of 3,750 lactating Holstein cows (1,563 primiparous cows and 2,187 multiparous cows) from a commercial dairy farm in Slovakia calving from January 2020 until July 2021 were enrolled on an ongoing basis. Activity data were recorded from d 7 until d 60 postpartum. Within this observational period, cows were classified into 3 categories: (1) no estrus event (Estrus0), (2) 1 estrus event (Estrus1), or (3) 2 or more estrus events (Estrus2+). Transition cow health was assessed by farm personnel within the first 30 d in milk (DIM) using standard operating procedures. Generalized linear mixed models were used to analyze continuous and categorical data. Cox proportional hazard models were used for time to event data. The overall prevalence of anestrus was 20.8%. Multiparous cows had a greater risk for anestrus compared with primiparous cows [odds ratio (OR) = 1.4]. Cows with stillbirth (OR = 1.76), retained placenta (OR = 2.19), puerperal metritis (OR = 1.48), or subclinical ketosis (OR = 1.51) had a greater risk for anestrus. In addition, cows calving in summer (OR = 0.82), autumn (OR = 0.38), or winter (OR = 0.56) had a higher incidence of anestrus than cows calving in spring. Estrous expression from d 7 until d 60 postpartum was associated with estrous duration (DU) and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest DU at first postpartum AI (9.4 ± 0.18 h) compared with cows in Estrus1 (10.5 ± 0.13 h) and Estrus2+ (11.4 ± 0.12 h). Cows in Estrus2+ had a longer DU at first postpartum AI compared with cows in Estrus1. For Estrus0, Estrus1, and Estrus2+ cows, pregnancy per AI at first service was 42.5%, 50.9%, and 55.4%, respectively. Estrous expression from d 7 until d 60 postpartum was associated with time to first AI and time to pregnancy. Compared with Estrus0 cows, Estrus1 [hazard ratio (HR) = 1.43] and Estrus2+ cows (HR = 1.62) had an increased hazard of being inseminated within 100 DIM. Compared with Estrus2+, Estrus1 cows had a reduced hazard of being inseminated within 100 DIM (HR = 0.89). Compared with Estrus0 cows, Estrus1 (HR = 1.24) and Estrus2+ cows (HR = 1.46) had an increased hazard of becoming pregnant within 200 DIM. Median DIM to pregnancy were 121, 96, and 92 for Estrus0, Estrus1, and Estrus2+ cows, respectively. In conclusion, cows with transition cow disorders (i.e., stillbirth, retained placenta, puerperal metritis, or subclinical ketosis) had a greater chance for anestrus compared with healthy cows. Cows in Estrus0 had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity from d 7 until d 60.     

Effect of lameness on ovarian activity in postpartum holstein cows

A longitudinal study was conducted to examine the relationship between lameness and delayed ovarian cyclicity during the first 60 d postpartum and days to first luteal activity during the first 300 d postpartum in Holstein cows. Two hundred thirty-eight cows from a 600-cow dairy that calved during a 12-mo period were used. Cows were classified into 1 of 6 categories of lameness during the first 35 d postpartum using a locomotion scoring system. Cows were blood-sampled weekly for detection of plasma progesterone concentrations during the first 300 d postpartum. Cows with delayed resumption of ovarian cyclicity were defined as those with progesterone concentrations consistently <1 ng/mL during the first 60 d postpartum. The null hypothesis that risk of delayed cyclicity is the same in cows classified as nonlame, moderately lame, or lame (after adjusting for potential modifying or confounding effects of loss of body condition and other variables related with delayed cyclicity) was tested using logistic regression. Analysis of results of the study reported here support the hypothesis that lameness is associated with delayed ovarian activity in Holstein cows during the early postpartum period. Cows classified as lame had 3.5 times greater odds of delayed cyclicity, compared with cows classified as nonlame. Attributable proportion analysis indicated that delayed ovarian cyclicity in lame cows would be reduced by 71%, if lameness had been prevented.     

Relationship of plasma nonesterified fatty acids and walking activity in postpartum dairy cows

To survive and produce milk, postpartum dairy cows use their reserves through lipolysis. If the negative energy balance is severe, nonesterified fatty acids (NEFA) are formed that can impair several physiological processes. A pilot study suggested that increased walking activity after calving may be related to a reduced serum concentration of NEFA. The objective of this study was to determine the relationship between plasma concentrations of NEFA and walking activity in dairy cattle during the postpartum period. Data were collected from 33 multiparous Holstein-Friesian dairy cows. Walking activities were quantified using pedometry, and blood samples were collected for determination of NEFA. Results of this study indicated that a negative relationship existed between walking activity and plasma NEFA concentrations in postpartum dairy cows.     

Technical note: Comparison of rectal and vaginal temperatures in lactating dairy cows

A method commonly used to identify illness in dairy cows is measuring body temperatures with a rectal thermometer, but vaginal measures are becoming common in research. The primary objective of this study was to validate vaginal measures of body temperature by comparing them with rectal temperatures. Data loggers used to collect vaginal temperatures can be programmed to collect many readings per day, providing an opportunity to interpret effects of health in relation to diurnal differences in temperatures. Thus, a secondary objective was to compare the diurnal pattern in body temperatures for cows with and without retained placenta (RP). Body temperature was monitored for 8 d in 29 cows that had recently calved (enrolled 2 d after calving; 7 of these cows were diagnosed with RP) and in 13 cows in peak lactation (98 ± 8 d in milk). Rectal temperatures were taken at 0630, 0930, 1230, 1530, 1830, and 2130 h (±30 min) with a digital thermometer for 8 d consecutively. During the same period, vaginal temperatures were measured every 10 min with a microprocessor-controlled data logger attached to a modified vaginal controlled internal drug release insert. Values from the vaginal loggers were averaged over 1 h and paired with the corresponding rectal temperature. There was a relationship between rectal and vaginal temperatures for fresh cows (n = 1,393; r = 0.81) and for peak-lactation cows (n = 556; r = 0.46). Cows with RP had higher body temperatures (39.2 ± 0.01) compared with healthy cows (39.1 ± 0.01). Body temperature was higher at night, and lower between 0800 to 1000 h for healthy cows (39.0 ± 0.02) and between 1100 to 1300 h for RP cows (39.1 ± 0.02). In summary, vaginal temperatures were associated with rectal measures, and provided the advantage of capturing dirurnal changes in body temperature.     

Genetic analysis of postpartum measures of luteal activity in dairy cows

The objective of this study was to estimate genetic parameters for measures of luteal activity during the first 60 d postpartum. Analyses were made with different sampling intervals to investigate the possibility of combining progesterone measurement with routinely performed milk recording. Progesterone level in milk as an indicator of female fertility when selecting sires in a progeny-testing scheme was also examined. Data were collected from 1996 to 1999, and comprised 1,212 lactations from 1,080 British Holstein-Friesian cows at 8 commercial dairy farms in the United Kingdom. Milk samples for progesterone analysis were collected thrice weekly. Mixed linear animal models were used to analyze the data. Heritability for the percentage of samples with luteal activity during the first 60 d postpartum (PLA) was 0.30 and decreased with more infrequent sampling to 0.25, 0.20, and 0.14 for weekly, twice-monthly, and monthly sampling, respectively. Measures of PLA had a high negative genetic correlation with prolonged anovulation (−0.53 for monthly sampling, < −0.87 otherwise) and a moderate positive genetic correlation with persistent corpus luteum in the first estrus cycle (>0.65 if at least twice-monthly sampling). Genetic correlations with interval from calving to commencement of luteal activity were close to −1 for all PLA measurements and the selection index calculations showed that monthly progesterone sampling could be used with high accuracy (0.80 with 50 daughters per bull) to predict breeding values for commencement of luteal activity. Progesterone analysis at the time of regular milk recording could thereby be used to select for an early interval from calving to commencement of luteal activity and, at the same time, a decreased frequency of prolonged anovulation during the postpartum period.     

Effects of synchronization treatments on ovarian follicular dynamics, corpus luteum growth, and circulating steroid hormone concentrations in lactating dairy cows

Lactating dairy cows (n=57) ≥45 d postpartum at first service were enrolled in a randomized complete block design study to evaluate treatments to synchronize estrus and ovulation. At 10 d before artificial insemination (AI), animals were randomly assigned to 1 of 3 treatments: (1) d -10 GnRH (GnRH1; 10 μg of buserelin, i.m.) and controlled internal drug release insert [CIDR, 1.38 g of progesterone (P4)]; d -3 PGF(2α) (PGF; 25 mg of dinoprost, i.m.); d -2 CIDR out; and AI at observed estrus (CIDR_OBS); (2) same as CIDR_OBS, but GnRH (GnRH2) 36 h after CIDR out and timed AI (TAI) 18 h later (CIDR_TAI); or (3) same as CIDR_TAI, but no CIDR (Ovsynch). Transrectal ultrasound was used to assess follicle size before ovulation and on d 4, 8, and 15 after the presumptive day of estrus (d 0) to measure the corpus luteum (CL). Blood samples were collected to determine concentrations of estradiol (E2; d -10, -9, -3, -2, -1, and 0) and P4 (d -10, -9, -2, -1, 0, 1, 4, 6, 8, 11, and 15). No treatment differences were observed in either circulating concentrations of P4 or the ovulatory response to GnRH1 at the onset of synchronization treatments. Circulating concentrations of P4 were greater for CIDR_OBS and CIDR_TAI compared with Ovsynch at 24 h after CIDR insertion (5.34 and 4.98 vs. 1.75 ng/mL) and immediately before CIDR removal (1.65 and 1.48 vs. 0.40 ng/mL). Peak circulating concentrations of E2 were greater for CIDR_OBS compared with Ovsynch (3.85 vs. 2.39 pg/mL), but CIDR_TAI (2.82 pg/mL) did not differ from either CIDR_OBS or Ovsynch. The interval from PGF injection to peak circulating E2 did not differ between CIDR_TAI and Ovsynch (52.1 vs. 49.8 h). Both CIDR_TAI and Ovsynch, however, had shorter intervals from PGF injection to peak circulating E2 concentrations compared with CIDR_OBS (67.8 h). The diameter of the dominant follicle before ovulation was greater for CIDR_OBS compared with Ovsynch (18.5 vs. 16.0 mm) but CIDR_TAI (17.1 mm) did not differ from either of the other treatments. The mean interval from PGF to ovulation was longer for CIDR_OBS (100.0 h) compared with CIDR_TAI and Ovsynch (84.4 and 83.2 h, respectively). Use of CIDR_OBS resulted in increased preovulatory follicle size and greater circulating concentrations of E2 due to a longer period of preovulatory follicle growth. Progesterone supplementation during synchronization and GnRH on the day before TAI affected ovulatory follicle size, and periovulatory circulating concentrations of P4 and E2. No differences, however, in postovulatory P4 or luteal volume profiles were observed.     

Graduate Student Literature Review: Effects of human chorionic gonadotropin on follicular and luteal dynamics and fertility in cattle

Circulating progesterone concentrations during the growth of the ovulatory follicle and early embryo development have been positively associated with embryo quality and survival and pregnancy success. As a potent luteotropic agent with LH-like activity, human chorionic gonadotropin (hCG) has been tested in different studies to improve pregnancy outcomes by increasing circulating progesterone concentrations during the growth of the ovulatory follicle or early embryonic development. Nevertheless, hCG has produced inconsistent, contradictory, and intriguing results. Furthermore, recent research indicates that hCG, when used before artificial insemination, may affect physiological events necessary for the ovulation of a viable oocyte. In addition, the use of hCG-inducing accessory corpus luteum during the estrous cycle seems to disturb luteolysis and follicle and luteal dynamics during the estrous cycle. This literature review discusses past and current research exploring the effects of hCG on the estrous cycle characteristics and pregnancy per artificial insemination and embryo transfer.     

Postpartum ovarian activity in multiparous Holstein cows treated with bovine somatotropin and fed n-3 fatty acids in early lactation

Multiparous cows (n = 59) were blocked by expected calving date and previous 305-d mature-equivalent milk yield and assigned randomly to a 2 × 2 factorial design to determine the effects of bovine somatotropin (bST; Posilac, Monsanto Animal Agricultural Group, St. Louis, MO) and dietary fat on ovarian activity during the first 90 d in milk (DIM). Diets that included whole, high-oil sunflower seeds [SS; 10% of dietary dry matter; rich in linoleic acid (18:2)] or a mixture of Alifet-High Energy and Alifet-Repro [AF; Alifet USA, Cincinnati, OH; 3.5 and 1.5% of dietary dry matter, respectively; protected source of linolenic (18:3), eicosapentaenoic, and docosahexaenoic fatty acids] were provided from calving. Diets were isocaloric at equal intakes, but AF provided more net energy for lactation at actual intakes (1.54 vs. 1.66 Mcal/kg of dry matter). Cows received 0 or 500 mg of bST (N, Y) every 10 d from 12 to 70 DIM and at 14-d intervals from 70 to 280 DIM. Breeding was initiated after 90 DIM. Follicular dynamics, luteal growth and development (15 to 90 DIM), and plasma progesterone concentrations (1 to 90 DIM) were evaluated (3 times per week). Days to first ovulation (33.6 ± 1.4) and incidence of anovulation at 45 or 70 DIM did not differ among treatments. Interovulatory intervals were similar among treatments (22.1 ± 0.9 d). Incidence of estrous cycles with 2 follicular waves was greater for SSY (71.0%) and AFN (80.0%) than for other groups, but more 3-wave cycles occurred with AFY (83.3%). Growth rate of the ovulatory follicle was greater for AF than SS (1.9 vs. 2.2 ± 0.11 mm/d) and diameter of ovulatory follicles was larger for AFN than the other treatments (17.9 vs. 15.7 ± 0.7 mm). Area under the progesterone curve was reduced for SSY (63.2, 48.1, 55.5, and 61.4 ± 5.1 ng·d/mL for SSN, SSY, AFN, and AFY, respectively). The number of class 1 (3 to 5 mm) follicles was decreased and the number of class 2 (6 to 9 mm) follicles was increased by bST. The number of class 2 follicles was reduced by AF. Initiation of bST administration at 12 DIM and dietary n-3 fatty acids altered ovarian activity during the first 90 DIM and could benefit reproductive performance. Dietary n-3 fatty acids interacted with bST administration in early lactation to increase the incidence of estrous cycles with 3 follicular waves. Although these changes could benefit reproductive performance, evaluation with a larger number of cows is needed to determine if these alterations improve fertility.     

Effects of a dietary supplement enriched in palmitoleic acid on fatty acid composition of follicular fluid,granulosa cell metabolism,and oocyte developmental capacity in early lactation dairy cows

In high-yielding dairy cows, some fertility traits can be influenced by the fatty acid (FA) composition of the follicular fluid during early lactation. The first objective of the current study was to evaluate the potential of dietary supplements enriched in specific FA to influence the FA composition of follicular fluid lipid classes in early lactation dairy cows. The second objective was to determine the influence of the resulting follicular fluid FA composition on the folliculogenesis, lipid and energy metabolism of granulosa cells, as well as oocyte quality and embryo development. Twenty Holstein multiparous cows in late gestation were randomly assigned to 200 g/d of FA supplements enriched in (1) palmitic acid (control treatment; 82% 16:0; PA) in the rumen or (2) palmitoleic acid (sea buckthorn oil; 27% cis-9 16:1, 28% 16:0, 22% cis-9 18:1, and 11% cis-9,cis-12 18:2; SBT) in the abomasum. The treatment period ranged from 20 ± 5 d precalving to 67 ± 2 d postcalving. Cumulus-oocyte complexes, granulosa cells, and follicular fluid were recovered from 2 sequential sessions of ovum pick-up (OPU-1 and OPU-2) at 46 and 67 ± 2 d postcalving (mean ± standard deviation). On the same days, blood samples were collected. Milk performance was recorded, and feed and milk samples were collected from d 8 to 10 ± 3 (onset of lactation), d 35 to 37 ± 2 (before OPU-1), and d 63 to 65 ± 2 (before OPU-2). Treatments did not affect milk yield or fat concentration throughout the experimental trial. Compared with PA, SBT increased the cis-9 16:1 concentration in milk fat, in plasma esterified lipid classes (phospholipids, cholesterol esters, and triacylglycerols), and in follicular fluid phospholipids and cholesterol esters at OPU-1. Abundance of mRNA for stearoyl-CoA desaturase 1 and 5, and perilipin 2 in granulosa cells was not different between treatments, but an increase in the level of stearoyl-CoA desaturase 5 was observed between the 2 OPU periods. Treatments did not affect oocyte quality and developmental capacity or embryo lipid metabolism when cultivated in vitro. These results suggest that limited modifications in the FA composition of the oocyte microenvironment via dietary lipid supplements enriched in specific FA had no major effects on granulosa cell metabolism and oocyte developmental capacity in early lactation cows.     

Inducing ovulation early postpartum influences uterine health and fertility in dairy cows

The objective of the current study was to evaluate the effect of GnRH early postpartum on induction of ovulation, uterine health, and fertility in dairy cows. Holstein cows without a corpus luteum (CL) at 17 ± 3 DIM were assigned randomly to receive i.m. GnRH (n = 245) at 17 ± 3 and 20 ± 3 DIM or remain as controls (n = 245). Ovaries were scanned by ultrasonography twice weekly totaling 4 examinations. Ovulation was characterized by the appearance of a CL ≥20 mm at any ultrasound or CL <20 mm in 2 consecutive examinations. Clinical and cytological endometritis were diagnosed at 35 DIM. Compared with control, GnRH increased ovulation up to 3.5 d after the last treatment (78.7 vs. 45.0%) and did not affect the prevalence of clinical endometritis (23.9 vs. 18.6%) or cytological endometritis (30.9 vs. 32.8%). Prevalence of clinical endometritis increased in cows that had calving problems (32.6 vs. 15.9%) and metritis (40.6 vs. 15.8%). Metritis increased prevalence of cytological endometritis (50.7 vs. 23.5%). Treatment with GnRH did not affect pregnancy per artificial insemination at 32 (37.6 vs. 38.6%) or 74 d after artificial insemination (35.0 vs. 31.5%), but reduced pregnancy loss (6.8 vs. 18.1%). No overall effect of GnRH treatment on hazard of pregnancy was observed; however, an interaction between GnRH treatment and ovulation showed that GnRH-treated cows that ovulated had increased hazard of pregnancy by 300 DIM compared with GnRH-treated and control cows that did not ovulate (hazard ratio = 2.0 and 1.3, respectively), but similar to control cows that ovulated (hazard ratio = 1.1). Gonadotropin-releasing hormone early postpartum induced ovulation without affecting uterine health, but failed to improve pregnancy per artificial insemination or time to pregnancy, although it reduced pregnancy loss.     

Association of activity and subsequent fertility of dairy cows after spontaneous estrus or timed artificial insemination

The objective of this observational study was to evaluate the association between increased physical activity at first artificial insemination (AI) and subsequent pregnancy per AI (P/AI) in lactating Holstein cows following spontaneous estrus or a timed AI (TAI) protocol. We also wanted to identify factors associated with the intensity of activity increase (PA) captured by automated activity monitors (AAM) and fertility. Two experiments were conducted, in which cows either were inseminated based on the alert of the AAM system (AAM cows) or received TAI following a 7-d Ovsynch protocol (TAI cows) if not inseminated within a farm-specific period after calving. Experiment 1 included 2,698 AI services from AAM cows and 1,042 AI services from TAI cows equipped with the Smarttag Neck (Nedap Livestock Management) from a dairy farm in Slovakia (farm 1). In the second experiment, 6,517 AI services from AAM cows and 1,226 AI services from TAI cows fitted with Heatime (Heatime Pro; SCR Engineers Ltd.) from 8 dairy farms in Germany (farms 2–9) were included. Pregnancy diagnosis was performed on a weekly basis by transrectal ultrasound (farms 1, 3, 7, 8) or by transrectal palpation (farms 2, 4–6, 9). Estrous intensity was represented by the peak value of the change in activity. In experiment 1, PA was categorized into low (x-factor 0–20) and high (x-factor 21–100) PA, and in experiment 2 into low (activity change = 35–89) and high (activity change = 90–100) PA. In TAI cows from both experiments, PA was additionally categorized into cows with no AAM alert. Data were analyzed separately for AAM and TAI cows using multinomial logistic regression models for PA in TAI cows and logistic regression models for PA in AAM cows and P/AI in both groups. In experiment 1, P/AI of AAM cows was greater for AI services performed with conventional frozen semen (57.6%) compared with sexed semen (47.2%), whereas type of semen only tended to be associated with P/AI in TAI cows (54.4% conventional frozen semen vs. 48.9% sexed semen). In experiment 2, P/AI was greater for fresh semen (AAM cows: 44.4% vs. TAI cows: 44.2%) compared with conventional frozen semen (AAM cows: 37.6% vs. TAI cows: 34.6%). In both experiments, pregnancy outcomes were associated with PA. In experiment 1, AAM cows with high PA (55.1%) had greater P/AI than cows with low PA (49.8%). Within TAI cows, cows with no alert (38.8%) had reduced P/AI compared with cows with low (54.2%) or high PA (61.8%). In experiment 2, AAM cows with high PA (45.8%) had greater P/AI compared with cows with low PA (36.4%). Timed AI cows with no alert (27.4%) had decreased P/AI compared with cows with low (41.1%) or high (50.8%) PA. The greatest risk factors for high PA were parity (experiment 1) and season of AI (except for TAI cows from experiment 1). We conclude that high PA at the time of AI is associated with greater odds of pregnancy for both AAM and TAI cows. In both experiments, about 2 thirds of AAM cows (experiment 1: 69.9% and experiment 2: 70.7%) reached high PA, whereas only approximately one-third or less of TAI cows (experiment 1: 37.3% and experiment 2: 23.6%) showed high PA. Although we observed similar results using 2 different AAM systems for the most part, risk factors for high PA might differ between farms and insemination type (i.e., AAM vs. TAI).