Effect of postpartum calcium supplementation on serum calcium and parathyroid hormone concentrations in multiparous Holstein cows.

Although postpartum Ca supplementation strategies are often employed to prevent subclinical hypocalcemia in dairy cows, these strategies have produced a mix of beneficial, neutral, and detrimental results when assessing milk yield and subsequent disease outcomes. Because the mechanisms underlying these differing results are unknown, our objectives were to determine how common postpartum Ca supplementation strategies affect blood Ca concentrations and parathyroid hormone (PTH). We conducted a randomized controlled trial with 74 multiparous dairy cows on a commercial dairy in central New York. Cows were assigned to 1 of 4 supplementation groups immediately after calving: (1) control (CON; no Ca supplementation, n = 15); (2) conventional oral Ca supplementation (BOL-C; 43 g of oral Ca bolus administered immediately after calving and 24 h later, n = 17); (3) delayed oral Ca supplementation (BOL-D; 43 g of oral Ca bolus administered 48 and 72 h after calving, n = 15); or (4) subcutaneous infusion (SQ; 500 mL of 23% Ca borogluconate infused subcutaneously once immediately after calving, n = 15). Blood samples were collected immediately after calving (0 h) and at 8, 16, 24, 32, 40, 48, 56, 64, 72, 80, 88, 96, 120, and 168 h postpartum for a total of 15 blood samples per cow. Cows were excluded if administered Ca, via any route, by farm employees or if they died or were sold within 96 h following parturition, which left 62 cows for analysis. Linear mixed models, accounting for repeated measures, were created to analyze changes in serum total Ca (tCa) and PTH over the first 168 h after parturition and assess differences between supplementation groups. Serum tCa and PTH concentrations were not different at the time of calving among supplementation groups. There was a supplementation group by hour postcalving interaction for mean tCa concentration in which SQ cows had reduced tCa concentrations from 32 to 64 h compared with CON cows, 32 to 96 h compared with BOL-C cows, and 40 to 64 h compared with BOL-D cows. Mean PTH concentration did not differ among supplementation groups across 168 h after enrollment and was 158.1 pmol/L (95% confidence interval [CI] = 148.2 to 168.0) for CON cows, 164.0 pmol/L (95% CI = 154.9 to 173.1) for BOL-C cows, 158.7 pmol/L (95% CI = 149.2 to 168.1) for BOL-D cows, and 153.2 pmol/L (95% CI = 143.6 to 162.8) for SQ cows. Our findings suggest that although serum tCa does not differ between cows that receive conventional or delayed oral Ca bolus supplementation at calving and cows that receive no supplemental Ca, subcutaneous infusion of Ca at calving reduces serum tCa for a substantial period between 32 and 64 h postsupplementation. However, as PTH concentrations did not differ among groups across 168 h postpartum, the mechanism by which tCa is reduced remains unclear.

Does knowledge of blood calcium concentration at 2 days postpartum affect decisions of calcium supplementation?

Delaying oral Ca supplementation might benefit cows with low blood Ca concentrations at 4 d in milk (DIM), a time when reduced blood total Ca (tCa) is associated with negative health and production outcomes. To implement a targeted approach to manage subclinical hypocalcemia (SCH) at the herd level, it is important to identify which cows benefit from supplemental Ca. Therefore, our objective was to determine if SCH diagnosis at 2 DIM could inform decisions of oral Ca supplementation at 2 and 3 DIM based on milk yield and 4 DIM blood Ca concentration. Data were analyzed from a previously conducted randomized controlled trial on multiparous cows (n = 518) from 4 farms in New York State. Cows were randomly assigned to 1 of 2 treatment groups at calving: (1) control (CON; no Ca supplementation, n = 259) or (2) bolus (BOL; 43 g of oral Ca administered at 2 and 3 DIM postcalving, n = 259). For each parity group (2, 3, 4+), we used generalized linear mixed models to identify serum tCa concentrations at 2 DIM that maximized the difference in milk yield to diagnose SCH. Cows were classified as normocalcemic (NC; parity 2 tCa >1.9 mmol/L, parity 3 tCa >1.87 mmol/L, n = 327; parity ≥4 had no defining threshold) or SCH (parity 2 tCa ≤1.9 mmol/L, parity 3 tCa ≤1.87 mmol/L, n = 58; parity ≥4 had no defining threshold). Parity 2 and 3 cows were further classified into 1 of 4 SCH-treatment groups (SCHTRT) based on 2 DIM SCH status and random treatment allocation: (1) NC-CON, n = 165, (2) SCH-CON, n = 28, (3) NC-BOL, n = 162, or (4) SCH-BOL, n = 30. Generalized linear mixed models were used to analyze the difference in milk yield for the first 10 wk of lactation and tCa at 4 DIM between SCHTRT groups with separate analyses performed for parities 2 and 3. Mean milk yield differed between SCHTRT groups for both parities. For parity 2, SCH-CON and SCH-BOL cows produced more milk than NC-CON and NC-BOL cows with SCH-CON producing 50.9 (95% confidence interval [CI] = 48.4, 53.4) kg/d, SCH-BOL 51.7 (49.1, 54.2) kg/d, NC-CON 47.5 (46.3, 48.7) kg/d, and NC-BOL 47.2 (45.8, 48.5) kg/d of milk. Milk yield was also different between SCHTRT groups for parity 3 with SCH-BOL cows producing more milk than NC-CON and NC-BOL cows. In parity 3, SCH-BOL cows produced 56.3 (95% CI = 53.1, 59.3) kg/d, SCH-CON 51.7 (48.6, 54.7) kg/d, NC-BOL 50.6 (49.0, 52.2) kg/d, and NC-CON 48.7 (46.9, 50.5) kg/d of milk. For both parities, SCH-CON and SCH-BOL cows had lower tCa at 2 DIM than NC-CON and NC-BOL cows. At 4 DIM, tCa concentrations were similar for all SCHTRT groups respective to parity. Our results suggest that although delayed Ca bolus administration does not improve blood Ca concentration when compared with controls, it does support increased milk production in parity 3 cows regardless of Ca status at 2 DIM. Thus, knowledge of blood Ca at 2 DIM should not affect decisions of Ca supplementation in this parity of cows.

Patterns of periparturient rumination and activity time in multiparous Holstein cows with and without dyscalcemia in early lactation.

Dyscalcemia, reduced blood Ca at 4 DIM, is associated with reduced milk production and reproduction and an increased risk of negative health events. Cowside testing of blood Ca to diagnose dyscalcemia is difficult, and alternative methods to identify dyscalcemia are needed. Our objectives were to explore differences in periparturient rumination and activity time between cows with and without dyscalcemia and use activity and rumination variables to identify dyscalcemia. We performed a retrospective cohort analysis on data collected from multiparous Holstein cows (n = 182) from 2 herds in NY. Cows were affixed with ear or neck loggers to record daily activity (arbitrary units (AU)/d, defined by manufacturer) and rumination (min/d) times. Daily activity and rumination times were collected from 14 d before calving until 14 DIM. No cows received supplemental calcium or experienced clinical hypocalcemia during the study period. A blood sample was collected at 4 DIM and analyzed for total calcium concentration, and cows were subsequently classified as dyscalcemic if total calcium ≤2.2 mmol/L (n = 57), or eucalcemic if total calcium >2.2 mmol/L (n = 125). Linear mixed models were used to analyze differences in pre- and postpartum activity and rumination times between calcemic groups. Logistic regression models were used to identify the probability of dyscalcemia from activity and rumination time variables from 0 to 4 DIM. Prepartum activity time was similar between eucalcemic and dyscalcemic cows (402.0 ± 10.4 AU/d and 395.1 ± 14.5 AU/d, respectively). Postpartum, eucalcemic cows had greater activity time than dyscalcemic cows (436.1 ± 10.7 vs. 407.8 ± 14.7 AU/d, respectively). Prepartum rumination time was similar between eucalcemic and dyscalcemic cows (512.6 ± 9.8 min/d and 504.2 ± 14 min/d, respectively). Postpartum, eucalcemic cows had greater rumination time than dyscalcemic cows (512.3 ± 10.5 min/d vs 480.5 ± 15 min/d, respectively). Logistic regression models yielded AUC values ranging from 0.71 to 0.79, sensitivities of 17.5 to 40.3%, specificities of 91.2 to 94.4%, accuracy of 70.3 to 77.0%, positive predictive values of 59.0 to 76.0%, and negative predictive values of 72.0 to 78.0%. Our findings suggest that differences exist in postpartum activity and rumination times between cows that experience dyscalcemia and those that remain eucalcemic. Utilizing activity and rumination time data in the immediate postpartum period shows utility in identifying cows with dyscalcemia, which could aid in management decisions that ameliorate the associated negative outcomes; however, further work is needed to optimize their capabilities.

Effects of timing of oral calcium administration on milk production in high-producing early-lactation Holstein cows.

Supplementation of oral Ca via blanket administration of an oral Ca bolus at 0 and 24 h after calving has shown limited success in increasing production and minimizing adverse health events. Recent evidence that reductions in blood Ca at 4 d in milk (DIM) are more closely associated with negative outcomes than hypocalcemia at 0 to 24 h postpartum might explain this lack of Ca bolus efficacy. Therefore, our primary objective was to explore the effect of delayed oral Ca bolus supplementation on milk production, with secondary objectives of exploring the effects on disease incidence and postpartum blood Ca dynamics. We conducted a randomized controlled trial on multiparous Holstein cows (n = 998) from 4 herds in New York. At calving, cows were randomly assigned to 1 of 3 treatment groups: (1) control, no supplemental Ca at or around parturition (CON; n = 343); (2) conventional bolus, an oral Ca bolus containing 43 g of Ca at calving and 24 h later (BOL-C; n = 330); or (3) delayed bolus, an oral Ca bolus containing 43 g of Ca at 48 and 72 h after calving (BOL-D; n = 325). We created generalized linear mixed models to analyze differences in milk yield for the first 10 wk of lactation and serum total Ca (tCa) at 1 and 4 DIM between treatment groups; multivariable Poisson regression models were used to analyze adverse event outcomes (metritis, displaced abomasum, herd removal, or a combination of one or more of the 3) in the first 30 DIM. Milk yield increased by week and was not affected by treatment. However, a treatment by parity group interaction for milk yield showed that BOL-D cows in the third parity produced more milk than third-parity BOL-C or CON cows (BOL-D = 52.0 kg/d, 95% confidence interval [50.6, 53.4] kg/d, BOL-C = 47.9 [46.3, 49.5] kg/d, CON = 49.8 [48.2, 51.2] kg/d). The incidence of adverse health events was similar between treatments (BOL-D = 3.7%, BOL-C = 3.7%, CON = 3.6%). Serum tCa was lower at 1 than 4 DIM, and we detected no difference in tCa between treatment groups. Our findings suggest that delaying oral Ca bolus supplementation has limited influence on blood Ca concentrations but may be beneficial to cohorts of cows as a targeted prophylactic supplement to support milk production.

A randomized field trial assessing the timing of postpartum calcium bolus administration on milk yield of multiparous Holstein cows.

Recent studies have shown that cows with subclinical hypocalcemia (SCH) at 4 d in milk (DIM), regardless of their blood Ca concentration before that time point, suffer from an increased early-lactation disease risk and reduced milk yield, whereas cows experiencing a transient reduction in blood Ca that regain normocalcemia by 4 DIM are at a reduced risk of disease and have greater milk yields. With a goal of improving outcomes for dyscalcemic cows with SCH at 4 DIM, our primary objective was to assess the effect of a herd-level oral Ca bolus strategy that delayed supplementation to 24 and 48 h postpartum on productive performance of multiparous Holstein cows. Our secondary objectives were to assess the effects of delayed Ca bolus supplementation on blood Ca concentration, disease incidence within 30 DIM, and pregnancy risk to first service. At calving, multiparous cows on a single commercial dairy farm in Iran were randomly assigned to 1 of 3 treatment groups: (1) control, no Ca bolus administration (CON; n = 95); (2) traditional bolus, one Ca bolus administered immediately following calving and a second Ca bolus administered 24 h after calving (TRD, n = 102); or (3) experimental bolus, one Ca bolus administered 24 h after calving with a second Ca bolus administered 48 h after calving (EXP, n = 99). Blood samples were collected at 0, 24, 48, 72, and 96 h, and 7 d after parturition, with sampling occurring before bolus administration for the TRD (0 and 24 h) and EXP (24 and 48 h) groups. A general linear mixed model was created to analyze the change milk yield over the first 4 monthly tests and serum Ca concentrations over 7 DIM. Given the lack of disease events diagnosed within 30 DIM, no statistical analysis was conducted for this outcome. The effect of treatment group on risk of pregnancy to first service was assessed using Poisson regression. The incidence of dyscalcemia within CON cows was 72%. We found no difference in mean monthly milk yield among treatment groups across the first 4 tests, with an average monthly production of 51.8 ± 8.8 kg/d for CON cows, 52.5 ± 8.7 kg/d for TRD cows, and 51.8 ± 8.7 kg/d for EXP cows. Mean blood Ca concentration also did not differ across 7 DIM among treatment groups and was 2.04 mmol/L [95% confidence interval (CI) = 2.00 to 2.07 mmol/L] for CON cows, 2.06 mmol/L (95% CI = 2.03 to 2.09 mmol/L) for TRD cows, and 2.09 mmol/L (95% CI = 2.05 to 2.12 mmol/L) for EXP cows. The risk of pregnancy to first service was numerically greater for CON than TRD and EXP cows but not statistically different; however, our study was underpowered for this outcome. Under the conditions of our study, our findings suggest that delaying oral Ca bolus supplementation to 24 and 48 h postpartum has no effect on milk production across the first 4 monthly tests.

Patterns of Fourier-transform infrared estimated milk constituents in early lactation Holstein cows on a single New York State dairy.

Cows undergo immense physiological stress to produce milk during early lactation. Monitoring early lactation milk through Fourier-transform infrared (FTIR) spectroscopy might offer an understanding of which cows transition successfully. Daily patterns of milk constituents in early lactation have yet to be reported continuously, and the study objective was to initially describe these patterns for cows of varying parity groups from 3 through 10 d postpartum, piloted on a single dairy. We enrolled 1,024 Holstein cows from a commercial dairy farm in Cayuga County, New York, in an observational study, with a total of 306 parity 1 cows, 274 parity 2 cows, and 444 parity ≥3 cows. Cows were sampled once daily, Monday through Friday, via proportional milk samplers, and milk was stored at 4°C until analysis using FTIR. Estimated constituents included anhydrous lactose, true protein, and fat (g/100 g of milk); relative % (rel%) of total fatty acids (FA) and concentration (g/100 g of milk) of de novo, mixed, and preformed FA; individual fatty acids C16:0, C18:0, and C18:1 cis-9 (g/100 g of milk); milk urea nitrogen (MUN; mg/100 g of milk); and milk acetone (mACE), milk ß-hydroxybutyrate (mBHB), and milk-predicted blood nonesterified fatty acids (mpbNEFA) (all expressed in mmol/L). Differences between parity groups were assessed using repeated-measures ANOVA. Milk yield per milking differed over time between 3 and 10 DIM and averaged 8.7, 13.3, and 13.3 kg for parity 1, 2, and ≥3 cows, respectively. Parity differences were found for % anhydrous lactose, % fat, and preformed FA (g/100 g of milk). Parity differed across DIM for % true protein, de novo FA (rel% and g/100 g of milk), mixed FA (rel% and g/100 g of milk), preformed FA rel%, C16:0, C18:0, C18:1 cis-9, MUN, mACE, mBHB, and mpbNEFA. Parity 1 cows had less true protein and greater fat percentages than parity 2 and ≥3 cows (% true protein: 3.52, 3.76, 3.81; % fat: 5.55, 4.69, 4.95, for parity 1, 2, ≥3, respectively). De novo and mixed FA rel% were reduced and preformed FA rel% were increased in primiparous compared with parity 2 and ≥3 cows. The increase in preformed FA rel% in primiparous cows agreed with milk markers of energy deficit, such that mpbNEFA, mBHB, and mACE were greatest in parity 1 cows followed by parity ≥3 cows, with parity 2 cows having the lowest concentrations. When measuring milk constituents with FTIR, these results suggest it is critical to account for parity for the majority of estimated milk constituents. We acknowledge the limitation that this study was conducted on a single farm; however, if FTIR technology is to be used as a method of identifying cows maladapted to lactation, understanding variations in early lactation milk constituents is a crucial first step in the practical adoption of this technology.

Blood calcium as a prognostic indicator of success after surgical correction of left displaced abomasum.

Subclinical hypocalcemia, a reduction in blood calcium concentrations without apparent clinical signs of milk fever, occurs in 25 to 50% of early-postpartum dairy cows. Research has shown that these cows are at an increased risk of additional early-lactation diseases, including left displaced abomasum (LDA), compared with their normocalcemic counterparts. However, no work has assessed the association of total calcium (tCa) concentration at the time of LDA correction with subsequent milk yield and survival within the herd. Pending future development of an economical and accurate on-farm test for hypocalcemia, the ability to assess LDA prognosis after surgical correction based on precorrection blood tCa concentration is of interest to practitioners. Therefore, our objective was to determine the association of blood tCa concentration at the time of LDA surgical correction with milk yield and herd removal in the 60 d after correction. Nine bovine practitioners, located in New York State, were involved in sample collection from 17 herds for this prospective cohort study. Immediately following LDA diagnosis and before surgical correction, blood samples were collected from 152 dairy cows within the first 30 d in milk for serum tCa determination. The association of tCa with milk yield (n = 110) and herd removal (n = 127) in the first 60 d after LDA surgical correction was analyzed using repeated-measures ANOVA and a generalized linear mixed model, respectively, controlling for parity and days in milk at LDA surgical correction and the random effect of herd or practitioner. We found no evidence to support an association of tCa at time of LDA diagnosis with subsequent milk yield, and when cows were grouped by median tCa into low and high groups (tCa ≤2.1 and >2.1 mmol/L, respectively), subsequent milk production was similar (39.4 ± 1.6 and 40.1 ± 1.4 kg/d for low and high, respectively). Furthermore, we found no evidence to support an association of tCa at time of LDA diagnosis with the odds of herd removal (odds ratio = 0.6; 95% confidence interval = 0.4-1.7). Therefore, our study provided no clear evidence to support the association of tCa at time of LDA diagnosis and correction with either milk yield or herd removal during the 8 wk after correction. However, it did demonstrate low to extremely low tCa concentration in cows with LDA. Whether calcium supplementation would improve health and production outcomes in these cows is unclear and worth investigating further.

Differential effects of a single dose of oral calcium based on postpartum plasma calcium concentration in Holstein cows.

Our objectives were to determine (1) the effect of a single dose of an oral Ca bolus within 24 h after parturition on plasma Ca concentration, (2) the response of primiparous (PP) and multiparous (MP) cows to this supplementation strategy, and (3) differential responses based on plasma Ca at enrollment. For objective 1, cows from 1 commercial dairy in New York State were enrolled within 19 h after parturition (mean ± standard deviation = 8.3 ± 5.3 h) and randomized within parity group (first, second, and ≥third) to control [CON (n = 25); no placebo] or a single dose bolus treatment [BOL (n = 25); 3 oral Ca boluses supplying 54 to 64 g of Ca]. Plasma Ca was measured repeatedly between 1 and 24 h following treatment. For objectives 2 and 3, cows on 6 commercial farms in New York State were assigned to treatment as for objective 1 (CON, n = 1,973; BOL, n = 1,976). Herd records for health, reproduction, and Dairy Herd Improvement Association test day milk production were collected. Mixed effect multivariable models were developed using repeated measures ANOVA, Poisson regression, or proportional hazard models. Objective 2 analyses considered treatment with periparturient risk factors, whereas objective 3 analyses also considered Ca status. No difference was observed for plasma Ca between 1 and 24 h after treatment. Primiparous cows assigned to BOL calving at >712 d old had decreased risk of one or more health disorders [≤30 d in milk; risk ratio (RR) = 0.65, 95% confidence interval (CI) = 0.51 to 0.84] and those with body condition score >3.5 responded to BOL with increased milk production (CON = 31.7 ± 1.1, BOL = 35.1 ± 1.1 kg/d), as did those with days carried calf >277 (CON = 31.9 ± 1.0, BOL = 34.7 ± 1.0 kg/d). Reduced risk of one or more health disorders was observed in parity ≥3 (RR = 0.85, 95% CI = 0.81 to 0.89) and MP cows with body condition score >3.5 (retained placenta; RR = 0.70, 95% CI = 0.58 to 0.84) or that were lame (displaced abomasum; RR = 0.49, 95% CI = 0.32 to 0.75). Differential responses for PP cows by Ca status were minimal. For MP cows with low plasma Ca, BOL decreased risk of additional Ca treatment (≤1.8 mmol/L; RR = 0.57, 95% CI = 0.40 to 0.80) as well as risk of one or more health disorders (≤2.15 mmol/L; RR = 0.90, 95% CI = 0.85 to 0.95). Supplementation with a single oral dose of Ca could be targeted to periparturient risk groups for improved health. Calcium status did not differentiate responses of PP cows, but MP cows with low Ca at parturition had improved health status when supplemented.

A stochastic estimate of the economic impact of oral calcium supplementation in postparturient dairy cows.

The objective was to develop stochastic models to estimate the economic impact in the first 30 d in milk of oral calcium supplementation to multiparous postparturient dairy cows using 4 different strategies: (1) supplementation of cows with a high previous lactation mature-equivalent milk yield, (2) supplementation of lame cows, (3) supplementation of both cows that have a high previous lactation mature-equivalent milk yield and cows that are lame, and (4) supplementation of all cows. Data from current literature were used to model input variables associated with the costs and risks related to milk production, postparturient disease, and culling. The mean net herd impact per 1,000 calvings for each of the 4 supplementation strategies was $4,425, $5,812, $8,313, and $3,065, respectively. Postpartum supplementation of multiparous lame cows had the highest return on investment at 6.5 to 1, followed by supplementation of multiparous high milk yield and lame cows, multiparous high milk yield cows only, and supplementation of all multiparous postpartum cows with returns of 1.8 to 1, 1.1 to 1, and 0.3 to 1, respectively. A herd's average milk yield at first test had the highest influence on the net impact of oral calcium supplementation to all multiparous cows and accounted for 30% of the variation, followed by the decrease in risk of health events in lame cows given oral calcium at 22%, a herd's prevalence of lameness at calving at 13%, and the price of milk at 10%. Each of the remaining stochastic variables contributed to less than 5% of the variation in net herd financial impact of oral calcium administration. Whereas supplementation of all postpartum multiparous cows returned a positive net herd impact approximately 80% of the time, if a herd was willing to devote time to mature-equivalent milk yield calculations and locomotion scoring, supplementation of this subpopulation of postpartum cows with oral calcium was estimated to have a positive economic impact in all iterations. Depending on the supplementation strategy chosen and baseline milk yield and immediate postpartum lameness prevalence in a herd, a herd with 1,000 calvings per year can expect to see an average net impact ranging from approximately $3,000 to $8,000 after postpartum supplementation of oral calcium in multiparous animals.

Effects of chromium propionate supplementation during the periparturient period and early lactation on metabolism, performance, and cytological endometritis in dairy cows.

Multiparous Holstein cows (n=61) were used to determine the effects of chromium propionate (Cr-Pro) supplementation during the periparturient period and early lactation on metabolism, performance, and the incidence of cytological endometritis (CE). After a 1-wk preliminary period, cows were assigned randomly to 1 of 2 treatments from 21 d before expected calving through 63 d postpartum: (1) control (n=31) and (2) Cr-Pro (n=30) administered by daily topdress at a rate of 8 mg/d of Cr. A tendency was detected for increased dry matter intake (DMI) during the prepartum period for cows fed Cr-Pro. Moreover, cows fed Cr-Pro tended to have lower plasma concentrations of nonesterified fatty acids during the prepartum period. However, effects of Cr-Pro supplementation on postpartum DMI and milk yield were not significant. Cows fed Cr-Pro tended to have higher urea N concentrations in milk. An interaction of treatment and day existed during the postpartum period, such that cows fed Cr-Pro had lower plasma glucose concentrations within the first day postpartum compared with controls. Plasma haptoglobin concentration was not affected by treatment during the postpartum period. Blood neutrophil glycogen concentrations were not affected by treatment when sampled at either 7 d postpartum or on one day between 40 and 60 d (48 d ± 0.44 standard error) postpartum. Evaluation of endometrial cytology by low volume lavage at 7 d postpartum (first lavage) and on one day between 40 and 60 d (second lavage) postpartum revealed that cows fed Cr-Pro tended to have a higher percentage of neutrophils at first lavage and decreased incidence of CE as assessed at second lavage. In conclusion, supplementation with Cr-Pro resulted in trends for increased DMI and lower plasma nonesterified fatty acids prepartum. Postpartum production and energy metabolism were not affected by treatment; however, Cr-Pro supplementation tended to affect the postpartum influx of neutrophils into the uterus and decreased the incidence of CE, suggesting positive effects of Cr-Pro supplementation on uterine health.