Milk Progesterone Concentrations Research Articles

Pilot study investigating estrus length and estrus behavior in Norwegian Red cattle on a commercial dairy farm.

Norwegian Red has been shown to have high levels of estrus behavior under experimental conditions. However, the estrus behaviors of Norwegian Red cows have not been studied under commercial conditions. A herd of 89 Norwegian Red cows housed in free stalls on concrete, slatted floors, were continuously video monitored for 21 days. Ovarian cyclicity was confirmed in a final study sample group (n = 18) using milk progesterone concentrations. All mounting and standing activities were recorded, and the duration of mount estrus, standing estrus and the differences between these; prestand and poststand, were determined. The cycle stages metestrus, diestrus and proestrus were estimated based on the starting time and ending time of mount estrus. All cows in the final study sample group exhibited the primary estrus sign, 'standing to be mounted' during estrus. Two (11%), eleven (61%) and six (33%) cows exhibited the behavior 'standing to be mounted' during metestrus, diestrus and proestrus, respectively. The number of mounts initiated by individual cows was higher during mount and stand estrus than during the rest of the estrous cycle. This study reports a median duration of mount estrus and stand estrus of 21.0 h (interquartile range (IQR) 15.0 to 27.3) and 14.3 h (IQR 12.0 to 18.8), respectively. The median counts per hour of all mount behaviors were 8.6 (IQR 5.6 to 11.3), 1.51 (IQR 0.3 to 3.8) and 1.7 (IQR 0.8 to 6.0) for standing estrus, prestand and poststand, respectively. This study shows that under commercial conditions the Norwegian Red cow displays a high level of mount and stand activity associated with estrus.

Milk fatty acids as indicators of delayed commencement of luteal activity in dairy cows in early lactation.

Excessive mobilization of adipose reserves due to severe negative energy balance in early lactation may be detrimental to dairy cow fertility at individual and herd level. Reproductive efficiency is one of the main factors influencing herd profitability and a strategy for early detection and management of cows with delayed resumption of cyclicity will result higher conception rate, decreased proportion of cows with extended lactation, fewer inseminations per conception and lower culling rates due to reproductive disorders. Using two groups of dairy cows (Holstein n=37, Swedish Red breed [SRB] n=49), we investigated potential differences between cows with different commencement of luteal activity (CLA) and the feasibility of using milk fatty acids (MFAs) as predictors of delayed CLA. Milk samples for progesterone analysis were collected twice weekly during the first six weeks in milk. The concentrations of the MFAs C14:0, C16:0, C18:0 and C18:1 cis-9 in milk (g/100 g milk) and in milk fat (g/100 g fat) were analysed by Fourier transform infrared spectroscopy and individual MFA profiles were calculated by weeks in milk. Commencement of luteal activity was defined as the first day with milk progesterone concentrations >3ng/ml at two successive measurements. The study population was categorized as early (n=42) or late (n=44) CLA, using the median value of 21 DIM as the cut-off. Analysis of the data revealed that CLA was correlated with the proportion of some specific MFAs, where cows with delayed CLA had lower IGF-1 (92.9 ± 7.9 vs. 114.1 ± 7.9ng/ml; p=.05) and C14:0 levels (10.4 ± 0.2 vs. 11.5 ± 0.2g/100 fat; p < .01) and higher C18:0 (9.6 ± 0.2 vs. 8.9 ± 0.6g/100 fat; p < .01) and C18:1 cis-9 levels (24.9 ± 0.4 vs. 23.5 ± 0.4g/100 fat; p < .05). Delayed CLA (mean 34 days) was predictable for approximately 80% of cows based on C18:0 or C18:1 cis-9 concentrations in week 2 postpartum. Overall, MFAs (C18:0 and C18:1 cis-9) as biomarkers were better indicators than beta-hydroxybutyrate or non-esterified fatty acids in early detection of cows with delayed or normal CLA. The MFA concentrations in milk samples from cows in early lactation can thus be used as a non-invasive method to identify cows at risk of delayed CLA, acting as potential biomarkers for future reproductive performance.

APPLICATION OF PROGESTERONE PROFILES IN THE ASSESSMENT OF POST ARTIFICIAL INSEMINATION FERTILITY IN CATTLE

The present study was conducted to investigate success of AI in relation to progesterone concentrations with respect to the timing of inseminations or heat detection in the field. Milk samples were collected for progesterone assay from 149 crossbred cows subjected to AI. The milk progesterone concentrations were estimated by RIA to access the reproductive status of the cows. On the day of AI, progesterone concentrations were less than 1.0 nmol/L (Low) in 51% samples and more than 3.00 nmol/L (High) in 18%. Rest 31 % samples showed intermediate progesterone (1.01 to 2.99 nmol/L) suggesting that these cows were nor in proper heat at the time of AI. On the basis of two samples (day 0 and day 10-12 post AI), 50% cows had low progesterone concentrations on the day of AI and high in the second sample, indicating that these cows were inseminated during estrus and ovulated subsequently. Whereas, 12% cows had high progesterone in both the samples suggesting that at the time of AI these cows were either in the luteal phase or had luteal cyst. Remaining 37% cow had intermediate progesterone concentrations on both the occasions suggesting that they were having either developing or regressing or malfunctioning corpora lutea or partial luteal cyst. Perusal of observations on progesterone concentrations showed that 18% of inseminations were done during luteal phase and another 30% were done at some inappropriate time of heat, thus suggesting that high proportion (48%) of cows under field conditions were inseminated while not in proper heat.

Feeding behavior in relation to ovarian cyclicity in cows with no or a short dry period

The objective of this study was to investigate relations between feeding behavior (FB) variables focusing on intake of the basal ration during 3 wk prepartum until 3 wk postpartum, and postpartum onset of luteal activity (OLA) in dairy cows subjected to no (0-d) or a short (30-d) dry period (DP). Feeding behavior of the basal ration intake (concentrate intake excluded) was continuously recorded by computerized feeders for 123 dairy cows and analyzed from 3 wk prepartum to 3 wk postpartum. Cows were subjected to a DP length: 0-d DP (n = 81), or 30-d DP (n = 42). Milk progesterone concentration was determined 3 times per week until 100 DIM to determine OLA, which was classified as early OLA (<21 DIM) and late OLA (≥21 DIM). Relations between FB and OLA class were analyzed using mixed models. During 3 wk prepartum, FB differed between parity class (parity 2 or ≥3 after calving). Cows with a 30-d DP prepartum, regardless of their OLA class, had more visits per meal, higher meal duration, total meal time, total daily feeding time, and lower feed intake and feeding rate compared with cows with 0-d DP. During the first 2 d postpartum, cows with OLA <21 had more visits per day and visits per meal compared with cows with OLA ≥21. During the first 3 wk postpartum, cows within the 30-d DP group with OLA <21 had greater meal size, feed intake, and feeding rate compared with cows with OLA ≥21 of this group. Cows within the 0-d DP group with OLA <21 had higher meal duration, total daily meal time, total daily feeding time, and meal size, but a lower feeding rate compared with cows with OLA ≥21 of this group. Concluding from the current study, cows with OLA <21 had a postpartum FB that reflected a cow with faster recovery from parturition and better adaptation to onset of lactation compared with OLA ≥21. No or a short DP affected prepartum FB, suggesting that prepartum management affects FB. This study provides evidence of different FB between cows with different timing of resumption of ovarian cyclicity.

Study of progesterone level during first trimester of pregnancy in cows.

Measurement of progesterone is an indirect method for pregnancy diagnosis in many livestock species including cattle, buffaloes, sheep, and goats. Conception extends the life of the corpus luteum (CL) by preventing the luteolytic mechanism from being triggered, thus prolonging and maintaining its functional characteristics, ensuring continued high progesterone levels (Spencer, et.al., 1995). (Shemesh et al., 1973) proposed that the difference in peripheral plasma progesterone levels between pregnant and non pregnant cows, 19 days after insemination, can form the basis for a very early pregnancy test. (Laing and Heap 1972) first documented this in milk to diagnose cows in early pregnancy Progesterone maintains the uterine endometrium in a state which supports embryonic development, implantation, and foetoplacental development. Progesterone concentrations vary with the stage of the estrous cycle which makes it one of the most commonly studied reproductive hormones in bovine ruminants for pregnancy detection and ovarian activity (Kaneko,et.al.,1994). Studies in the bovine estrous cycle indicate that the milk or serum progesterone concentrations reach a maximum value 13-14 days after estrus, and if the animal is pregnant, these continue to remain elevated up to day 21 after fertilization (Parkinson, et al.,1994) . These high levels of progesterone in serum or milk between days 18 and 24 after insemination form the basis of establishment of pregnancy in cattle (Sasser, and Ruder,1987; Shemesh,1973). Interferon-???? exerts its antiluteolytic effect by inhibiting the endometrial expression of oxytocin receptors, through which oxytocin stimulates pulsatile PGF2???? release (Wolf,2003). Although low progesterone concentrations at 18 to 24 days after breeding can accurately predict non pregnancy, high progesterone concentrations during this period are not the specific indicators of pregnancy due to variations among cows in duration of the estrous cycle as well as the incidence of early or late embryonic mortality. The advantages of progesterone assay for pregnancy diagnosis include noninvasive collection of milk sample and the feasibility to conduct the test on the farm using commercial cow-side milk progesterone test kits [Pennington,1985 ; Gowan,1982 ; Wimpy,et.al.,1986), though the sensitivity gets compromised to some extent with these assay kits.&#x0D; Key Word: progesterone, pregnancy, cows

Pointing Out Some Issues Regarding Reproduction Management in Murciano-Granadina Goats.

Simple SummaryThe hypothesis of this experiment proposes that it could be possible to identify pregnant goats through maximum progesterone milk levels at any time in the pregnancy, and that there is an optimal moment to apply a lactation inhibitor to dry off lactating goats. The maximum progesterone concentration in milk varied depending on the season of the year, and those concentrations were similar for pregnant and non-pregnant goats, but significantly higher in the case of gestating goats with four foetuses, for which it would be possible to distinguish the pregnancy. The milk yield of goats at mating does not affect fertility until a value of at least 3250 mL/day. If using lactation inhibitors, their application up to the 10th week post-mating would be optimal for drying off lactating goats.Two of the most important problems in high-yielding dairy goat farms are early and accurate pregnancy diagnosis and the appropriate dry off of lactating does before the next kidding. The hypothesis posits that it could be possible to identify pregnant does through maximum progesterone milk levels at any time during the pregnancy, and that there is an optimal time to apply a lactation inhibitor to help dry off lactating does. Therefore, 114 Murciano-Granadina breed goats were used, from which 74 goats were inseminated at week 20 of lactation and samples of milk from pregnant and non-pregnant goats were taken at two-week intervals. The average maximum progesterone milk levels were higher outside the natural breeding season (40° latitude) than in the breeding season (11.6 ± 1.13 vs. 8.6 ± 1.02 ng/mL), although the levels from pregnant and non-pregnant goats were similar (10.85 ± 1.3 vs. 9.74 ± 1.6 ng/mL), except in the case of pregnancy with four foetuses (12.5 ± 1.3 ng/mL). Milk yield at mating does not affect fertility until a value of at least 3250 mL/day. Pregnancy started to affect milk yield up to the +7th week and was 59.9% lower in the +10th week after mating, so the use of lactation inhibitors could be more effective from this latter week. In conclusion, the results show that it is not possible to detect gestation in goats reliably through the maximum concentration of progesterone in milk at any time during lactation, except in the case of goats gestating four foetuses, that the milk yield of goats at mating does not affect fertility until a value of at least 3250 mL/day, and that from the 10th week post-mating, the application of lactation inhibitors would be optimal.

Relationship between in-line milk progesterone before and after artificial insemination and fertility outcomes in dairy cows.

The aim of this study was to evaluate the possibility to predict outcomes of artificial insemi- nation (AI) in dairy cows based on in-line milk progesterone (P4) concentration. The research was carried out on the herd of loose housing 245 dairy cows of 2-4 lactations, with average milk yielding 11.000 kg per cow. Milk sampling, measuring, and recording of milk P4 concentration was carried out using the Herd Navigator (HN). The grouping was performed according to the following three indices: the first by reproductive condition - pregnant or not pregnant after AI, the second by P4 concentration from day 20 before AI to day 20 after AI, and the third by P4 concentration at AI time. There was a significant difference in P4 concentration in the group of pregnant cows from day 15 to day 9 before AI, and it was by 18.3% higher com- pared to that in the group of non-pregnant cows in the said period (p⟨0.01). The milk P4 concen- trations began to differ mostly from day 10 after AI. At that time, the average P4 concentration in the group of pregnant dairy cows was by 36.8% higher compared to that in the group of non-pregnant cows (p⟨0.01). A statistically significant difference between the ratio of the cows with high, medium, and low P4 concentration on days 20-16 before AI (p⟨0.01) was determined. The highest number of cows with up to 2-3 ng/ml P4 concentration became pregnant at the AI time. In-line milk P4 records captured on day 10-15 before AI can be used to predict the proper for reproduction period. By P4 concentrations on day10 after AI, the ratio of pregnant cows in herd can be assessed.

Study of criteria for selection and evaluation of donor cows in their preparation for embryo transplantation

In the present studies, we studied and tested various criteria for evaluating donors before carrying out fetal transplantation. The main objectives of the study were: to study the prevalence of gynecological pathology in cows, which are potential candidates for embryo transplantation, to determine the relationship between the concentration of carotene and the level of progesterone as a marker of ovarian activity. It was found that the greatest number of complications during childbirth were recorded during the period of fetal excretion, which were observed in 72% of animals, and in the subsequent stage of labor, which were observed in 68% of animals. In the postpartum period, inflammation of the genital organs was most often manifested in 23.6% and uterine subinvolution 13.89%. Further, in the postpartum period, the concentration of progesterone in milk in cows was investigated and the functional activity of the corpus luteum was determined. For the study, a group of 32 cows was formed whose genital involution had ended. The study of progesterone was carried out in dynamics, twice on the 35th and 45th days after calving. The study found that in 71.86% of cows the amount of progesterone was less than 5 ng / ml, which indicates a low activity of the corpus luteum. In 28.13% of animals the concentration of progesterone on the 35th day was higher than 5.67 ng / ml and averaged 8.22±3.83 ng / ml and significantly decreased to 1.86 ± 1.69 ng / ml. The number of cows with a consistently low P4 concentration in two measurements was 56.25% of the total number of animals in the group. Thus, it was found 12.5% with a reduced activity of the corpus luteum in the luteal phase. Also, in experimental animals, a biochemical study of blood serum was carried out and a pronounced relationship with the concentration of carotene in the blood was determined.

Cow Milk Progesterone Concentration Assessment during Milking Using Near-infrared Spectroscopy

Cow Milk Progesterone Concentration Assessment during Milking Using Near-infrared Spectroscopy

Postpartal resumption of ovarian activity in dairy cows: Implications for herbage-based feeding systems in Switzerland.

Milk production in Switzerland is mainly based on herbage feeding with little input of concentrates. The present study investigated the effects of a solely herbage-based diet with (C) and without concentrate (nC) supplementation on luteal activity, milk production and metabolic status in 23 multiparous Holstein dairy cows with early or delayed resumption of ovarian cyclicity post partum (pp). Cows were retrospectively assigned either to a group with early (until d 25 pp, EOV) or delayed resumption of ovarian activity (> d 30 pp, DOV), resulting in four subgroups depending on concentrate feeding: DOV-C, DOV-nC, EOV-C, EOV-nC. Milk progesterone (P4) concentration was measured every 3 d, and different metabolites were analyzed in weekly blood samples. Resumption of ovarian activity was detected between d 19 and 25 pp in EOV, and between d 30 and 60 pp in DOV. In DOV-C, resumption of cyclicity tended to start earlier (38.3 ± 1.7 d pp) compared to DOV-nC (45.2 ± 6.3 d pp; P = 0.10). Independent of grouping, the ovarian activity occurred later in cows with greater plasma β-hydroxybutyrate (BHB) concentrations (P 1 ng/mL did not differ between groups (P > 0.05), but milk P4 peaks during the experiment were higher in EOV compared to DOV (P .

The influence of lameness on several automatic milking system variablesand reproductive performance indicators in dairy cows

There is an increased interest in using automatic milking systems (AMS) to indirectly assess the welfare of dairy cows, but knowledge on analyzing the association between lameness, milk yield characteristics, and reproductive performance in cows is still insufficient. The main aims of this study were to evaluate the influence of lameness on several AMS variables and reproduc- tive performance indicators during the early stage of lactation and estrus in Lithuanian Black and White dairy cows, as well as to assess the associations between lameness, productivity and repro- ductive efficiency. A total of 418 milking cows (50.3±1.2 d postpartum) without any apparent reproductive disorder were monitored for hoof health status. Cows were assigned to two groups on the basis of visual locomotion scoring: "non-lame"cows (group 1; 74.20%) and cows presen- ting "lameness" (lame cows) (group 2; 25.80%). Productive and milking performances of dairy cows were recorded from 50 to 100 days in milk (DIM) and 1 day after the first estrus. The lameness was predominantly localized on the hind feet (79.60%) and less frequently - on the front feet (20.40%; p⟨0.001). Furthermore, the lameness had a tendency to decrease milk production (4.24%; p⟨0.05) and increase the diffe- rence in milk yield between rear and front quarters of the udder (1.20%; p⟨0.05). The frequency of milking (5.19%) was lower in lame cows (p⟨0.05). The lame cows during estrus showed a more pronounced decrement in milk yield and milking frequency (p⟨0.05), and also higher milk progesterone concentration values (1.55-1.76 time's; p⟨0.001), and an increasing number of inseminations (11.69%; p⟨0.05) were observed. The results highlighted that analysis of data from AMS programs can be a successful tool for reducing risk factors related to the effective management of reproductive performance and hoof health of dairy cows.

Dynamic Changes in Progesterone Concentration in Cows' Milk Determined by the At-Line Milk Analysis System Herd NavigatorTM.

Simple SummaryAccording to the literature, the at-line progesterone monitoring system Herd NavigatorTM (Lattec I/S, Hillerød, Denmark) was used in combination with a DeLaval milking robot (DeLaval Inc., Tumba, Sweden). It works automatically and provides real-time physiological information about lactating dairy cows. For making farm-management decisions, it is not only a novel tool for scientific research, but also a mechanism for improving productivity, food safety, animal well-being, the environment, and the public perception of the dairy industry. It has been hypothesized that the progesterone concentration determined by the at-line milk analysis system and changes in its dynamics correlate with the parity, reproductive status, and milk yield of cows. The aim of the current study was to evaluate relative at-line milk progesterone (mP4) dynamic changes, according to the parity and status of reproduction, and to estimate the relationship with productivity in dairy cows. Frequent automated mP4 sampling can help identify characteristics of mP4 dynamic changes associated with successful pregnancies, pregnancy losses, and potential differences in mP4 dynamics among parity groups, which have not been studied previously.The aim of the current instant study was to evaluate relative at-line milk progesterone dynamic changes according to parity and status of reproduction and to estimate the relationship with productivity in dairy cows by at-line milk analysis system Herd NavigatorTM. According to the progesterone assay, experimental animals were divided into three periods: postpartum, after insemination, and pregnancy. In the first stage of the postpartum period, progesterone levels in milk were monitored every 5 days. This period of reproductive cycle recovery was followed for 30 days (days 0–29). The second stage of the postpartum period (30–65 days) lasted until cows were inseminated. In the period (0–45 days) after cow insemination, progesterone levels were distributed according to whether or not cows became pregnant. For milk progesterone detection, the fully automated real-time progesterone analyzer Herd NavigatorTM (Lattec I/S, Hillerød, Denmark) was used in combination with a DeLaval milking robot (DeLaval Inc., Tumba, Sweden). We found that an at-line progesterone concentration is related to different parities, reproductive statuses, and milk yield of cows: the 12.88% higher concentration of progesterone in milk was evaluated in primiparous cows. The average milk yield in non-pregnant primiparous cows was 4.64% higher, and in non-pregnant multiparous cows 6.87% higher than in pregnant cows. Pregnancy success in cows can be predicted 11–15 days after insemination, when a significant increase in progesterone is observed in the group of pregnant cows.

Consequences of Transition Treatments on Fertility and Associated Metabolic Status for Dairy Cows in Early Lactation.

Simple SummaryShortening or omitting the dry period improves energy balance and metabolic status, but reduces milk production and increases the risk of body fattening of cows in the subsequent lactation. Reducing the postpartum dietary energy level in order to match the lower milk yield after 0-d dry period could prevent body fattening. Earlier, reducing postpartum dietary energy level for cows after 0-d dry period reduced days open in the subsequent lactation, which may indicate improved underlying fertility. This study investigated effects of reducing dietary energy level from week 4 postpartum onwards for cows after 0-d dry period on fertility variables and associated metabolic status. Reducing the postpartum dietary energy level in cows with 0-d dry period reduced the interval from calving to onset of luteal activity in cows of parity ≥ 3, compared with a standard dietary energy level or a 30-d dry period. Fewer days open was related to fewer services per conception, fewer days to onset of luteal activity, higher percentage of ovarian cycles of normal length (18–24 d), and improved energy balance in weeks 1–7 of lactation. In conclusion, reducing a postpartum dietary energy level to match lower milk yield after 0-d dry period improved fertility in cows of parity ≥ 3, but not in cows of parity 2.This study aimed to (1) investigate effects of reducing postpartum dietary energy level for cows after a 0-d dry period (DP) on resumption of ovarian cyclicity and reproductive performance, (2) relate days open with other reproductive measures, and (3) relate onset of luteal activity (OLA) and days open with metabolic status in early lactation. Holstein-Friesian dairy cows were randomly assigned to 1 of 3 transition treatments: no DP and low postpartum dietary energy level from 22 days in milk( DIM )onwards (0-d DP (LOW)) (n = 42), no DP and standard postpartum dietary energy level (0-d DP (STD)) (n = 43), and a short DP and standard postpartum dietary energy level (30-d DP (STD)) (n = 43). Milk progesterone concentration was determined three times per week until 100 DIM. Plasma metabolite and hormone concentrations were measured weekly until week 7 postpartum. Reducing postpartum dietary energy level in older cows (parity ≥ 3) after no DP and 22 DIM did not affect milk production but prevented a positive energy balance and shortened the interval from calving to OLA. In addition, services per pregnancy and days open were reduced in cows of parity ≥ 3 on 0-d DP (LOW), compared with cows of parity ≥ 3 with 0-d DP (STD), but not in cows of parity 2.

Reliable identification of pregnant dairy cows by double milk progesterone analysis

In view of unsatisfactory conception rates commonly encountered in high producing dairy operations reliable ways of early pregnancy detection are of practical relevance. By analyzing the milk progesterone concentration 3 weeks after insemination non-pregnant cows can be reliably detected, whereas the accuracy of detecting pregnant cows is known to be less reliable. A field study was conducted on 25 North German dairy farms. Weekly milk samples were analyzed for progesterone content from the first to the seventh week after insemination. The accuracy of prediction of an existing pregnancy, based on a high progesterone level 3 weeks after insemination, was 80.1% as compared to palpation per rectum which served as gold standard. The prediction accuracy of samples taken at any time between 19 and 23 days after insemination was virtually similar to that of samples taken punctually on day 20 or 21. The accuracy of prediction could be substantially improved by analyzing an additional milk sample. When collecting the additional sample 1, 2, 3 or 4 weeks after the 3 week-sample, prediction accuracy was increased to 90.3%, 91.6%, 94.7% and 97.0%, respectively. Double milk progesterone analysis of samples drawn 3 and 7 weeks after insemination provides a prediction accuracy of pregnant cows comparable to that of palpation per rectum, transvaginal ultrasonography or PAG determination. The capability to reliably detect open cows in time to have them re-inseminated at the first return-estrus and, by analyzing an additional milk sample, equally reliably detect existing pregnancies, yields the milk progesterone test a valuable management tool. It will become even more attractive with the anticipated availability of practicable on-farm milk progesterone tests and access to robotic sampling equipment.

Genome-wide association study of normal and atypical progesterone profiles in Holstein-Friesian dairy cows

Female fertility has a major role in dairy production and affects the profitability of dairy cattle. The genetic progress obtained by traditional selection can be slow because of the low heritability of classical fertility traits. Endocrine fertility traits based on progesterone concentration in milk have higher heritability and more directly reflect the cow's own reproductive physiology. The aim of our study was to identify genomic regions for 7 endocrine fertility traits in dairy cows by performing a genome-wide association study with 54,000 SNP. The next step was to fine-map targeted genomic regions with significant SNP using imputed sequences to identify potential candidate genes associated with the normal and atypical progesterone profiles. The association between a SNP and a phenotype was assessed by a single SNP analysis, using a linear mixed model that included a random polygenic effect. Phenotypes and genotypes were available for 1,126 primiparous and multiparous Holstein-Friesian cows from research herds in Ireland, the Netherlands, Sweden, and the United Kingdom. In total, 44 significant SNP associated with 7 endocrine fertility traits were identified on Bos taurus autosome (BTA) 1-4, 6, 8-9, 11-12, 14-17, 19, 21-24, and 29. Three chromosomes, BTA8, BTA17, and BTA23, were imputed from 54,000 SNP genotypes to the whole-genome sequence level with Beagle version 4.1. The fine-mapping identified several significant associations with delayed cyclicity, cessation of cyclicity, commencement of luteal activity, and inter-ovulatory interval. These associations may contribute to an index of markers for genetic improvement of fertility. Several potential candidate genes reported to affect reproduction were also identified in the targeted genomic regions. However, due to high linkage disequilibrium, it was not possible to identify putative causal genes or polymorphisms for any of the regions.

Technical note: Validation of an automated in-line milk progesterone analysis system to diagnose pregnancy in dairy cattle

The in-line milk analysis system (IMAS) is an automated biosensor technology that samples and quantifies milk progesterone concentrations (P4c) at frequent intervals starting early postpartum until pregnancy. The objective was to validate the use of pregnancy notifications (PregN) generated by an IMAS based on P4c profiles after artificial insemination (AI) to determine pregnancy and nonpregnancy status in dairy cows. Records of 1,821 AI events from 715 Holstein cows that had milk P4c (ng/mL) measured every 2.2 ± 1.9 d (mean ± standard deviation) between 24.5 ± 8.2 and 173.4 ± 49.3 d in milk through a real-time IMAS (Herd Navigator, DeLaval International, Tumba, Sweden) were evaluated. Based on variations in adjusted milk P4c (< vs. ≥ the 5.0 ng/mL threshold), the system determined the sampling frequency, onset and cessation of luteal phases, and pregnancy. If a luteal phase initiated (P4c increased to ≥5.0 ng/mL) after AI and remained uninterrupted, a PregN was generated starting at (mean ± standard deviation) 31.0 ± 4.3 d until 53.4 ± 7.9 d after AI, when sampling stopped, unless a decline in P4c (to <5.0 ng/mL) occurred indicating nonpregnancy and imminent estrus. The assessment of IMAS PregN at 4 weekly intervals was tested, and a confirmed calving occurrence between 262 and 296 d after AI, with no other subsequent AI recorded, was the gold standard for pregnancy. In total, 14.1 (256/1,821), 41.0 (746/1,821), and 50.7% (924/1,821) of AI events were followed by a decline in P4c before 19, 23, and 30 d after AI, respectively. Frequency of the last 3 sampling events preceding P4c decline was greater if P4c decline occurred between 18 and 25 d after AI (1.4 ± 0.5 samples per day) compared with before 17 or beyond 26 d after AI (1.0 ± 0.5 samples per day). At 30 ± 3 (27 to 33) d after AI, PregN occurred in 46.8% (853/1,821) of AI events, of which 15.2% (130/853) had a decline in P4c between 30 and 55 d after AI and 17.1% (146/853) was later confirmed nonpregnant based on the gold standard. A total of 40.7% (742/1,821) of AI events was confirmed pregnant by the gold standard, which was no different than the proportion of PregN at 51 ± 3 (48 to 54) d (40.9%; 744/1,821). At any time point between 27 and 54 d after AI, sensitivity and negative predictive values for PregN were greater than 95.0 and 96.0%, respectively, whereas specificity values were less than 90.0% for PregN before 40 d but greater than 94.0% for PregN beyond 41 d after AI. In conclusion, IMAS is able to diagnose pregnancy based on P4c profiles with high precision and determine early nonpregnancy based on the spontaneous cessation of the luteal phase. However, for accuracy greater than 95.0%, pregnancy declaration based on IMAS notifications alone should occur no earlier than 41 d after AI.

Using in-line milk progesterone data to characterize parameters of luteal activity and their association with fertility in Holstein cows

Our objectives were to characterize parameters of luteal activity based on milk progesterone concentration (P4c) data from before and after artificial insemination (AI) and to evaluate their potential association with fertility in Holstein cows. Records of AI events (n = 4,353) and of milk P4c (n = 158,961) obtained through an in-line milk analysis system (Herd Navigator, DeLaval International, Tumba, Sweden) from 1,891 lactations of 1,423 Holstein cows were evaluated. Milk P4c (ng/mL) were measured every 2.2 ± 1.9 d (mean ± standard deviation) between 23.6 ± 7.3 and 185.3 ± 56.7 d in milk. Variations in milk P4c of consecutive records were used to determine onset of luteal phase (increase in P4c from <5.0 to ≥5.0 ng/mL), luteal phase length (period, in days, of P4c ≥5.0 ng/mL), cessation of luteal phase (decline from ≥5.0 to <5.0 ng/mL, designated as P4c-decline), and pregnancy (AI followed by a luteal phase that remained uninterrupted until 50 d post-AI). The length of the luteal phase preceding AI, the highest P4c (P4c peak) during the luteal phase preceding AI, the lowest P4c preceding AI (P4c pre-AI) that followed a P4c-decline, and the interval between P4c-decline and AI were evaluated, as well as the interval between AI and onset of luteal phase, and P4c at early diestrus (4.5 ± 0.6 d post-AI), mid diestrus (10.0 ± 0.6 d post-AI), and late diestrus (14.1 ± 0.6 d post-AI). Data were analyzed using logistic regressions, and comparisons made based on quartiles and cut-points established by receiver operating characteristic curve analysis. Overall probability of pregnancy was 32.0%. Parameters associated with reduced probability of pregnancy (represented as percentage points decrease in the probability of pregnancy) were (1) luteal phase length >14.4 d (7.6% decrease), (2) P4c peak ≤24.7 ng/mL (4.5% decrease), (3) P4c pre-AI >0.5 ng/mL (5.5% decrease), (4) interval between P4c-decline and AI of >1.6 d (4.0% decrease), (5) interval between AI and onset of luteal phase of <7 or >11 d (9.3 and 12.1% decrease, respectively), and (6) P4c at early diestrus ≤0.7 or >3.5 ng/mL (15.2 and 6.7% decrease, respectively), (7) P4c at mid diestrus ≤12.4 ng/mL (12.5% decrease), and (8) P4c at late diestrus ≤22.7 ng/mL (9.7% decrease). The parameters of luteal activity associated with reduced probability of pregnancy established here could be used as benchmarks while developing recommendations to improve reproductive performance in herds using in-line milk progesterone monitoring.

Automated oestrus detection using multimetric behaviour recognition in seasonal-calving dairy cattle on pasture

AIM To evaluate the performance of a novel accelerometer-based oestrus detection system (ODS) for dairy cows on pasture, in comparison with measurement of concentrations of progesterone in milk, ultrasonographic examination of ovaries and farmer observations. METHODS Mixed-breed lactating dairy cows (n=109) in a commercial, seasonal-calving herd managed at pasture under typical farming conditions in Ireland, were fitted with oestrus detection collars 3 weeks prior to mating start date. The ODS performed multimetric analysis of eight different motion patterns to generate oestrus alerts. Data were collected during the artificial insemination period of 66 days, commencing on 16 April 2015. Transrectal ultrasonographic examinations of the reproductive tract and measurements of concentrations of progesterone in milk were used to confirm oestrus events. Visual observations by the farmer and the number of theoretically expected oestrus events were used to evaluate the number of false negative ODS alerts. The percentage of eligible cows that were detected in oestrus at least once (and were confirmed true positives) was calculated for the first 21, 42 and 63 days of the insemination period. RESULTS During the insemination period, the ODS generated 194 oestrus alerts and 140 (72.2%) were confirmed as true positives. Six confirmed oestrus events recognised by the farmer did not generate ODS alerts. The positive predictive value of the ODS was 72.2 (95% CI=65.3–78.4)%. To account for oestrus events not identified by the ODS or the farmer, four theoretical missed oestrus events were added to the false negatives. Estimated sensitivity of the automated ODS was 93.3 (95% CI=88.1–96.8)%. The proportion of eligible cows that were detected in oestrus during the first 21 days of the insemination period was 92/106 (86.8%), and during the first 42 and 63 days of the insemination period was 103/106 (97.2%) and 105/106 (99.1%), respectively. CONCLUSIONS and CLINICAL RELEVANCE The ODS under investigation was suitable for oestrus detection in dairy cows on pasture and showed a high sensitivity of oestrus detection. Multimetric analysis of behavioural data seems to be the superior approach to developing and improving ODS for dairy cows on pasture. Due to a high proportion of false positive alerts, its use as a stand-alone system for oestrus detection cannot be recommended. As it is the first time the system was investigated, testing on other farms would be necessary for further validation.

Association of production diseases with motor activity-sensing devices and milk progesterone concentrations in dairy cows

We have previously established that the efficiency of identifying oestrus with activity-sensing devices can be compromised by common production diseases; the present study was undertaken to determine how these diseases may affect device readings. A total of 67 Holstein-Friesian cows, >20 days postpartum, were equipped with activity-sensing neck collars and pedometers, and simultaneous milk progesterone profiles were also monitored twice a week. The influences of common production stressors on maximum activity and progesterone values were analysed. Approximately 30% potential oestrus events (low progesterone value between two high values) remained unrecognised by both activity methods, and progesterone values in these animals were higher on the potential day of oestrus when both activity methods did not detect an event (0.043 ± 0.004 versus 0.029 ± 0.004 ng/mL; P = 0.03). Data from a subset of 45 cows (two events each) were subjected to mixed models and multiple regression modelling to investigate associations with production diseases. Cow motor activity was lower in lame cows. Maximum progesterone concentrations prior to oestrus increased as time postpartum and body condition score (BCS) increased. There were also fewer days of low progesterone prior to oestrus associated with increases in BCS and maximum progesterone concentrations prior to oestrus. In conclusion, lameness was associated with lower activity values, but this suppression was insufficient to account for lowered oestrus detection efficiency of either device. However, associations were identified between production diseases and progesterone profiles.

The relationship between milk progesterone and milk traits during estrus in dairy Cows

The aim of this study was to evaluate relationship between milk progesterone concentration (P4) and milk traits at the start of estrus time and 12h after start of the estrus in dairy cows. The 96 milk samples of 48 Lithuanian dairy cows without reproduction disorders and 90–100 days after calving were evaluated. Cows were classified into two groups based on milk yield per day: less than 30 kg (n=20) and e”30 kg (n=28). Data were categorized by milk fat and protein content at the start estrus and 12h after start of estrus to evaluate relationship between P4 and milk traits examined. P4 at estrus time in dairy cows was significantly positively correlated with milk yield (P less than 0.001), whereas it was negatively correlated with milk protein (P less than 0.05-P less than 0.01) and fat at 12h after start of estrus. Dairy cows with F/P from 1.0 to 1.5 had the lowest P4 in milk. Results of the pregnancy in dairy cows were related with lower P4 and milk yield level (P less than 0.001), higher milk fat (P less than 0.05) and milk protein content (P less than 0.001). These cows had 1.90 times lower prevalence of the signs of subclinical ketosis (P less than 0.05) at estrus time when compared with non-pregnant cows. As a result, it was clearly demonstrated that P4 in dairy cows can help to evaluate and improve the reproductive properties of cows.