Intramammary infections in lactating Jersey cows: Prevalence of microbial organisms and association with milk somatic cell count and persistence of infection.

There are limited data available regarding pathogens causing intramammary infections (IMI) in Jersey cows. The objectives of this study were to characterize the prevalence of IMI caused by different microorganisms in lactating Jersey cattle and evaluate the associations among microbes and somatic cell count (SCC) and persistence of IMI. This prospective, observational, longitudinal study included lactating Jersey cows (n = 753) from 4 farms within a 415 km radius of Columbia, Missouri. Quarter foremilk samples were aseptically collected monthly for 3 consecutive months. Microorganisms were identified using aerobic milk culture and MALDI-TOF mass spectrometry. A commercial laboratory measured SCC using flow cytometry. Milk culture results were used to classify single microorganism infections as persistent (same microorganism species identified at first sampling and one other sampling) or nonpersistent infection. Mixed models were built to evaluate the associations between IMI status and SCC natural logarithm (lnSCC), as well as persistence and lnSCC. Overall, staphylococci were the most commonly isolated microorganisms among the 7,370 quarter-level milk samples collected. Median prevalence (using all 3 samplings) of specific microbes varied among farms; however, Staphylococcus chromogenes was a common species found at all farms. The most common microbial species that persisted were Staph. chromogenes, Staphylococcus aureus, Staphylococcus simulans, and Streptococcus uberis. Streptococcus dysgalactiae and Staph. aureus were the IMI associated with the most inflammation based on lnSCC. The small number of herds included in this study with the large variation in herd type limits the generalizability of the data. However, results of this study seem to be similar to those of previous studies in other breeds, suggesting management factors are more important than breed-specific differences when evaluating causes of IMI and associated subclinical mastitis.

Estimates of intra- and interclass correlation coefficients for rump touches and the number of steps during estrus in postpartum cows.

Estrus traits have economic value in dairy production systems and could be incorporated into genetic selection indices. In an effort to further understand selection responses, 2 studies were performed to estimate the intra- and interclass correlation coefficients for estrus traits. Holstein-Friesian cows (n = 1,197; study 1) across 5 pasture-based grazing dairy herds were fitted with a capacitive touch sensing (CTS) device on the rump (FlashMate, Farmshed Labs Limited, Hamilton, New Zealand). The daily number of rump touches were subjected to a peak detection program to objectively identify periods of increased rump touches above baseline (indicative of estrus). The number of times touched and the sum of the touch duration were used to compare farms and estimate the intraclass correlation (repeatability). For study 2, postpartum Holstein (n = 85) and Guernsey (n = 5) cows in a confinement-style dairy were used. Cows were fitted with an IceQube accelerometer (IceRobotics Ltd., Edinburgh, United Kingdom) to measure steps taken per hour and a CTS device was applied to both rumps. The interclass correlation for the number of rump touches and number of steps taken during estrus was calculated. Data collected from 5 herds (study 1) demonstrated a 2- to 3-fold difference between herds in the number of rump touches and total touch time during estrus. The intraclass correlation (repeatability; estimates of maximum heritability) for rump touches during estrus was 0.22. For study 2, the number of steps and the number of rump touches during estrus increased in a synchronous manner. The intraclass correlation (repeatability) for number of steps during estrus was 0.26. The interclass correlation (r) for the number of rump touches and the number of steps was 0.46 (R2 = 0.21). Based on the R2, at least 20% of the variation in the number of steps during estrus was explained by the number of touches to the rump of the cow. Selecting cows for the number of steps taken during estrus could increase the number of rump touches (mounts, chin rests, and so on, received from other cows) if a genetic correlation exists for the phenotypic correlation that we observed.

Short communication: Simultaneous measurements of estrus behavior and plasma concentrations of estradiol during estrus in lactating and nonlactating dairy cows.

Estrus is an important behavior that can potentially be subjected to genomic selection. Circulating estradiol concentrations at estrus may be a useful phenotype if the absolute concentrations of estradiol are associated with overt phenotypes for estrus (activity, rump touches, or both; e.g., mounts, chinrests) that can be easily observed. The objective was to measure plasma estradiol concentrations at estrus and associate these measurements with the increase in activity (steps per hour) and rump touches received at estrus. We also tested the effect of lactation on the estrus traits that we measured. Cows (n = 11 lactating and n = 9 nonlactating) were treated with PGF2α to synchronize estrus. A jugular vein was cannulated to collect blood every 2 h for plasma estradiol measurement. Plasma LH was measured during the periestrual period to determine the time of the LH surge. Cows were fitted with an accelerometer to measure activity (steps per hour) and a capacitive touch sensing device to measure the number of rump touches and total touch time. Plasma estradiol concentrations were poorly correlated with overt signs of estrus during the period leading up to maximum estrus activity. After peak estrus activity (when cows were going out of estrus and plasma estradiol concentrations were decreasing), a stronger correlation was detected between overt signs of estrus and plasma estradiol concentrations. Effective selection for improved estrus expression based on plasma estradiol concentrations will depend on whether the cow is coming into or going out of estrus at the time of blood sampling. An association existed between lactation and fewer number of hours in estrus when estrus was defined by an increase in activity (steps per hour). Lactating cows had a shorter interval from the onset of estrus to the LH surge, and the shorter interval to the LH surge may have reduced the period of elevated estradiol during estrus in the lactating cows. Understanding mechanisms that control the sensitivity of the cow to estradiol and making appropriate selection decisions based on these mechanisms will likely increase overt signs of estrus in dairy cows.

Ovarian function and the establishment and maintenance of pregnancy in dairy cows with and without evidence of postpartum uterine disease.

Uterine disease early postpartum reduces fertility during the breeding period. One potential mechanism involves the reduced functional capacity of the uterus to support pregnancy. A second potential mechanism involves damage to ovarian follicles associated with systemic inflammation. We categorized lactating Holstein cows into healthy (n = 63) and diseased (n = 39) uterus groups based on the percentage of polymorphonuclear neutrophils in the uterine lumen during the second and third month postpartum and evaluated the functionality of their ovaries and their capacity to establish and maintain pregnancy. Cows were enrolled in a timed artificial insemination protocol (Presynch Ovsynch) so that the first artificial insemination was approximately 75 d postpartum. Ovarian follicles and corpora lutea were counted and measured using transrectal ultrasound, ovulatory responses were assessed, and luteal phase progesterone concentrations were measured. Pregnancy was detected on d 18, 20, 22, 25, 32, and 45 through chemical (d 18 to 25) or ultrasonographic methods (d 32 and 45). The percentage of cows ovulating during the Presynch period; the number, diameter, and ovulatory capacity of follicles during the Ovsynch period; and plasma progesterone concentrations following ovulation were similar for healthy and diseased cows. The initial period of pregnancy establishment (d 18 to 22) appeared to be unaffected by disease because a similar percentage of healthy and diseased cows were pregnant during this period. Embryonic loss occurred in both healthy and diseased cows after d 22. Based on a relatively small number of pregnancies (n = 30 healthy and n = 17 diseased), the cumulative embryonic loss after d 22 was greater in diseased compared with healthy cows. In short, uterine disease as defined in this study did not affect cyclicity, ovarian follicular growth, or plasma progesterone concentrations. Percentages of healthy and diseased cows that were pregnant were similar from d 18 to 22 after artificial insemination. Greater embryonic loss was observed after d 22 in diseased compared with healthy cows, but this observation was based on a small number of pregnancies and should be studied further in larger trials with greater statistical power.

Effect of diets containing sulfate or chloride-based anionic salts, fed to grazing prepartum dairy cows, on concentrations of Ca in plasma, disease incidence and milk yield.

AIMS: To compare the effect of two partial mixed ration diets containing either sulfate or chloride-based anionic salts, fed to grazing prepartum dairy cows, on concentrations of Ca, Mg, P and beta-hydroxybutyrate (BHB) in plasma, disease incidence and milk yield during the early postpartum period. METHODS: The study was conducted on a seasonal-calving grazing dairy farm located in Benton (MO, USA). In August 2016, two groups of 200 cows were randomly assigned at 30 days before expected parturition to receive either a sulfate or chloride-based anionic diet, to achieve a dietary cation-anion difference (DCAD) of -25 mEq/kg DM. Cows were fed partial mixed rations to provide 50% of their DM intake, with 50% provided by grazing fescue pasture. Urine samples were collected from 20 cows from each group once a week for 5 weeks until calving for pH assessment. At parturition, 42 cows per group were selected at random and blood samples collected on Days 1, 2, 3 and 7 postpartum. The study continued until 100 cows per group calved. Health events and cumulative milk yield until 30 days postpartum were recorded. Concentrations of Ca, P, and Mg in plasma at Days 1, 2, 3, and 7, and BHB in plasma at Day 7 postpartum were assessed. RESULTS: Mean urine pH was lower in cows in the chloride than sulfate group (p≤0.06) on four of five sampling occasions. Mean concentrations of Ca in plasma on Day 1 were 2.00 (95% CI=1.94-2.05) and 1.91 (95% CI=1.88-1.97) mmol/L for the chloride and sulfate groups, respectively (p=0.15), and on Day 3 were 2.03 (95% CI=1.97-2.08) and 1.90 (95% CI=1.84-1.96) mmol/L, respectively (p=0.038). Concentrations of Mg, P and BHB in plasma and incidence of diseases postpartum were similar in both groups (p>0.05). There were no cases of clinical hypocalcaemia in either group. CONCLUSIONS AND CLINICAL RELEVANCE: Prepartum cows fed a partial mixed ration with a negative DCAD containing chloride-based anionic salts and free access to a fescue pasture had lower prepartum urine pH, and higher concentrations of total Ca in plasma during the first week postpartum, than cows receiving a partial mixed ration containing sulfate-based anionic salts.

Technical note: Evaluation of fine needle aspiration cytology for the diagnosis of fatty liver in dairy cattle.

Fatty liver is a common condition affecting dairy cattle during the periparturient period, characterized by a pathological accumulation of triglycerides (TG) in the hepatocytes. The objective of this study was to evaluate the diagnostic potential of fine needle aspiration cytology in fresh liver specimens using liver TG concentrations as a gold standard. Fifty-seven liver samples from Holstein cows were collected during processing at a slaughterhouse. Tissue and fine needle aspirate samples were obtained from the parietal upper portion of the caudate lobe. Two samples of liver tissue were collected with a 16 gauge × 15 cm biopsy needle for histological and TG concentration assessment. A third sample was collected for cytology using an 18 gauge × 5.08 cm needle. The contents of the needle were transferred to a glass slide, spread, and air-dried. Liver samples were assayed by colorimetry/fluorimetry to determine TG concentrations. Concentrations of TG <2% were considered normal. Histological and cytological evaluations were conducted by 2 different pathologists blind to the visual classification. Sensitivity (Se) and specificity (Sp) were calculated. Cytology had a Se and Sp of 73 and 85%, respectively. Histopathology had a Se and Sp of 45.9 and 100%, respectively. The likelihood of having higher scores for histopathology and cytology increased as a function of liver TG content (mg/g).

Predicting hyperketonemia by logistic and linear regression using test-day milk and performance variables in early-lactation Holstein and Jersey cows.

Although cowside testing strategies for diagnosing hyperketonemia (HYK) are available, many are labor intensive and costly, and some lack sufficient accuracy. Predicting milk ketone bodies by Fourier transform infrared spectrometry during routine milk sampling may offer a more practical monitoring strategy. The objectives of this study were to (1) develop linear and logistic regression models using all available test-day milk and performance variables for predicting HYK and (2) compare prediction methods (Fourier transform infrared milk ketone bodies, linear regression models, and logistic regression models) to determine which is the most predictive of HYK. Given the data available, a secondary objective was to evaluate differences in test-day milk and performance variables (continuous measurements) between Holsteins and Jerseys and between cows with or without HYK within breed. Blood samples were collected on the same day as milk sampling from 658 Holstein and 468 Jersey cows between 5 and 20 d in milk (DIM). Diagnosis of HYK was at a serum ß-hydroxybutyrate (BHB) concentration ≥1.2 mmol/L. Concentrations of milk BHB and acetone were predicted by Fourier transform infrared spectrometry (Foss Analytical, Hillerød, Denmark). Thresholds of milk BHB and acetone were tested for diagnostic accuracy, and logistic models were built from continuous variables to predict HYK in primiparous and multiparous cows within breed. Linear models were constructed from continuous variables for primiparous and multiparous cows within breed that were 5 to 11 DIM or 12 to 20 DIM. Milk ketone body thresholds diagnosed HYK with 64.0 to 92.9% accuracy in Holsteins and 59.1 to 86.6% accuracy in Jerseys. Logistic models predicted HYK with 82.6 to 97.3% accuracy. Internally cross-validated multiple linear regression models diagnosed HYK of Holstein cows with 97.8% accuracy for primiparous and 83.3% accuracy for multiparous cows. Accuracy of Jersey models was 81.3% in primiparous and 83.4% in multiparous cows. These results suggest that predicting serum BHB from continuous test-day milk and performance variables could serve as a valuable diagnostic tool for monitoring HYK in Holstein and Jersey herds.

Hot topic: 16S rRNA gene sequencing reveals the microbiome of the virgin and pregnant bovine uterus.

We tested the hypothesis that the uterus of virgin heifers and pregnant cows possessed a resident microbiome by 16S rRNA gene sequencing of the virgin and pregnant bovine uterus. The endometrium of 10 virgin heifers in estrus and the amniotic fluid, placentome, intercotyledonary placenta, cervical lumen, and external cervix surface (control) of 5 pregnant cows were sampled using aseptic techniques. The DNA was extracted, the V4 hypervariable region of the 16S rRNA gene was amplified, and amplicons were sequenced using Illumina MiSeq technology (Illumina Inc., San Diego, CA). Operational taxonomic units (OTU) were generated from the sequences using Qiime v1.8 software, and taxonomy was assigned using the Greengenes database. The effect of tissue on the microbial composition within the pregnant uterus was tested using univariate (mixed model) and multivariate (permutational multivariate ANOVA) procedures. Amplicons of 16S rRNA gene were generated in all samples, supporting the contention that the uterus of virgin heifers and pregnant cows contained a microbiome. On average, 53, 199, 380, 382, 525, and 13,589 reads annotated as 16, 35, 43, 63, 48, and 176 OTU in the placentome, virgin endometrium, amniotic fluid, cervical lumen, intercotyledonary placenta, and external surface of the cervix, respectively, were generated. The 3 most abundant phyla in the uterus of the virgin heifers and pregnant cows were Firmicutes, Bacteroidetes, and Proteobacteria, and they accounted for approximately 40, 35, and 10% of the sequences, respectively. Phyla abundance was similar between the tissues of the pregnant uterus. Principal component analysis, one-way PERMANOVA analysis of the Bray-Curtis similarity index, and mixed model analysis of the Shannon diversity index and Chao1 index demonstrated that the microbiome of the control tissue (external surface of the cervix) was significantly different from that of the amniotic fluid, intercotyledonary placenta, and placentome tissues. Interestingly, many bacterial species associated with postpartum uterine disease (i.e., Trueperella spp., Acinetobacter spp., Fusobacteria spp., Proteus spp., Prevotella spp., and Peptostreptococcus spp.) were also present in the uterus of virgin heifers and of pregnant cows. The presence of 16S rRNA gene sequence reads in the samples from the current study suggests that the uterine microbiome is established by the time a female reaches reproductive maturity, and that pregnancies are established and maintained in the presence of a uterine microbiome.

Technical note: Validation of a chemical pregnancy test in dairy cows that uses whole blood, shortened incubation times, and visual readout.

Chemical pregnancy testing is an alternative to traditional methods of pregnancy diagnosis (either manual palpation or ultrasound) in postpartum dairy cows and heifers. The objective was to validate a chemical pregnancy test that confers the advantages of using whole blood, rapid incubation times, and visual readout. Blood and milk samples were collected from Holstein dairy cows [n=320; 162±62 (mean ± SD) d in milk] on a confinement farm in northeast Missouri at 28 d after artificial insemination (AI). The samples were assayed for pregnancy-associated glycoproteins (PAG) by using a rapid visual test as well as traditional plasma- and milk-based tests. Transrectal ultrasonography diagnosis for pregnancy at 35 to 38 d after AI was the reference (gold) standard for all PAG tests. One hundred fifty-nine cows were diagnosed as pregnant by the reference standard (pregnancies per AI=49.7%). The tests were ELISA and either optical density (OD; measured with a microtiter plate reader; plasma, milk, and rapid visual tests) or visual readout (rapid visual test) were used to diagnose pregnancy. When OD was used, the percentage of pregnant cows classified correctly (sensitivity) for the plasma, milk, and rapid visual tests were 97±1, 96±2, and 95±1% (±SE), respectively. The sensitivity of the rapid visual test when assessed visually was 98±1%. The specificity (proportion of nonpregnant cows classified correctly) for the plasma, milk, and rapid visual was 94±2%, 94±2%, and 93±2% when an OD was used. When read visually, the specificity of the rapid visual test was lesser (85±3%) because some cows with faint visual signals yielded false positive diagnosis. The overall accuracy (proportion of pregnant and nonpregnant cows diagnosed correctly) was similar for all tests (plasma, milk, rapid visual OD, and rapid visual; 96±1, 95±1, 94±1, and 92±2%, respectively). In a second experiment, lactating Holstein cows (n=291) from 4 commercial confinement dairy farms in western Kentucky were tested 25 to 95 d after AI using the rapid visual test. The OD of the rapid visual test followed the known profile for PAG in circulation during the first trimester of pregnancy. The conclusion is that the rapid visual test has equal sensitivity and accuracy as existing PAG tests. A slightly lower specificity was found when the rapid visual test was read visually.

Short communication: Test for nonpregnancy in dairy cows based on plasma progesterone concentrations before and after timed artificial insemination.

Timed artificial insemination (AI) programs have increased reproductive efficiency in dairy herds. A low timed AI pregnancy per AI is partially explained by cows that fail to respond optimally to the series of treatments that are designed to synchronize ovulation for AI. We hypothesized that testing cows for plasma progesterone concentrations during a timed AI protocol could be used as an early diagnostic test for nonpregnancy. Lactating Holstein cows (n=160) in 2 confinement-style dairies were used. Cows were treated with Presynch Ovsynch 56 for timed AI. Concentrations of progesterone in plasma were measured at -3, 0, 7, and 25 d relative to timed AI. Progesterone data were analyzed and receiver operating characteristic curves were generated by using logistic regression. The area under the receiver operating curves for a progesterone test for nonpregnancy on d -3 (PGF2α), 0 (AI), 7, and 25 d relative to timed AI were 0.68, 0.52, 0.55, and 0.89, respectively. The cutpoints and sensitivity (respectively) for the progesterone test were 0.51ng/mL (lower=nonpregnant) and 28.2% for the day of PGF2α, 0.43ng/mL (greater=nonpregnant) and 17.9% for the day of AI, 1.82ng/mL (lower=nonpregnant) and 23.1% for 7 d after AI, and 2.67ng/mL (lower=nonpregnant) and 76.0% for 25 d after AI. The false positive rate was less than 5% for all tests. Analysis of a second data set from a published study gave approximately the same cutpoints and sensitivity. When both studies were combined, approximately 20% of nonpregnant cows could be identified with a single test that was done before or shortly after AI with a false positive rate of less than 5%. When 2 and 3 tests were applied sequentially, the sensitivity for identifying nonpregnant cows increased from 38.4 to 50.5%. The pregnancy per AI for those cows that met the established progesterone criteria was approximately 3 to 4 times greater than those that failed to meet the criteria. The conclusions were that cows destined to be nonpregnant after timed AI can be identified before or shortly after AI. Testing for nonpregnancy before or shortly after AI may have utility with respect to eliminating a nonproductive AI (cows identified before AI) or shortening the time to reinsemination (cows identified by 1 wk after AI).

Follicular populations and luteal function in dairy heifers treated with a controlled internal drug release insert for 14 days as a method to synchronize the estrous cycle before prostaglandin F2α treatment and artificial insemination.

Progesterone-containing controlled internal drug release (CIDR) inserts are used to synchronize the estrous cycle before PGF2α is administered for timed AI (14dCIDR-PGF2α program). The program, initially designed for beef cattle, was recently shown to be efficacious in dairy heifers. We hypothesized that the 14-d CIDR treatment would synchronize the estrous cycle in dairy heifers and result in a uniformly sized corpus luteum (CL) and largest follicle (LF) at the time of PGF2α treatment. Holstein (n=110) or Holstein × Guernsey (n=4) dairy heifers were assigned to 2 treatments: (1) 14dCIDR-PGF2α [CIDR in for 14 d, CIDR out for 16d, PGF2α and AI after observed estrus (n=57)] or (2) control [PGF2α and AI after observed estrus (n=57)]. Regardless of treatment, additional PGF2α injections were administered at 14-d intervals to heifers that were not seen in estrus. Ovarian ultrasonography and blood sampling were done on d 0 (CIDR administered), 14 (day CIDR removed), 19 (5d after CIDR removed), 30 (PGF2α administered), and 44 (second PGF2α dose administered to heifers that were not detected in estrus after the first PGF2α). Compared with control (untreated), more CIDR-treated heifers were categorized as having a small CL (≤ 9.9 mm) and large LF (15.0-19.9 mm) on d 14 (CIDR removal) and, as expected, a greater percentage of CIDR-treated heifers were in estrus during the 5d after the CIDR removal compared with control heifers (75.4 vs. 22.8%, respectively). On d 19, the CIDR-treated heifers had apparently ovulated based on disappearance of LF and appearance of small CL. On d 30 (PGF2α administration), 89% of 14dCIDR-PGF2α heifers had CL that were ≥ 20 mm in diameter compared with 55% for control. Presence of larger CL on d 30 was associated with greater concentrations of plasma progesterone in 14dCIDR-PGF2α compared with control (10.5 ± 0.5 vs. 5.0 ± 0.6 ng/mL, respectively). The percentages of heifers with LF in the smallest category (≤ 9.9 mm) tended to be less (5.3 vs. 16.6%) and the percentage of heifers with LF in the medium-size category (10.0 to 14.9 mm) tended to be greater (84.2 vs. 69.1%) for 14dCIDR-PGF2α versus control, respectively, on d 30. More heifers were detected in estrus within 5d after the first PGF2α (86.0 vs. 56.1%) and conception rate to AI using sexed semen tended to be greater (61.2% vs. 40.6%) for 14dCIDR-PGF2α compared with control (respectively). Treating dairy heifers with a CIDR for 14 d was an effective method to synchronize an estrous cycle before PGF2α was administered.

Hot topic: Comparison of sex-sorted and conventional semen within a fixed-time artificial insemination protocol designed for dairy heifers.

The objective was to compare pregnancy per AI (P/AI) with conventional (CON) or sex-sorted (SS) semen from a single sire within a fixed-time AI (FTAI) program designed for dairy heifers. Holstein heifers (n=240) were assigned to treatment (CON or SS) according to body weight and reproductive tract score. All heifers underwent FTAI by using the "Show-Me-Synch" protocol [controlled internal drug release (CIDR) insert from d 0 to 14 followed by PGF(2α) (25mg i.m.) 16d after insert removal (d 30) with GnRH (100 µg i.m.) and FTAI at 66 h after PGF(2α)]. A single professional technician performed the FTAI. Heifers were fitted with heat detection patches at PGF(2α) to characterize estrous response. Estrous response did not differ between CON (63/120; 53%) and SS (70/120; 58%) treatments. The CON heifers, however, achieved greater FTAI P/AI (82/120; 68%) compared with SS (45/120; 38%) heifers. The P/AI did not differ for CON heifers that exhibited or failed to exhibit estrus before FTAI [44/63 (70%) vs. 38/57(67%), respectively]. For SS heifers, however, those that exhibited estrus had greater P/AI compared with those that failed to exhibit estrus [32/70 (46%) vs. 13/50 (26%)]. Pregnancy per AI resulting from FTAI was greater for heifers that were inseminated with CON semen compared with those that received SS semen. The expression of estrus before FTAI did not affect P/AI when CON semen was used, whereas the P/AI with SS semen was greater for heifers detected in estrus. Further studies are required to develop strategies for using sex-sorted semen when inseminating heifers at predetermined fixed times on the basis of expression of estrus before FTAI.

Reproduction in grazing dairy cows treated with 14-day controlled internal drug release for presynchronization before timed artificial insemination compared with artificial insemination after observed estrus.

Progesterone-releasing (controlled internal drug release, CIDR) devices inserted for 14 d are used to presynchronize the estrous cycle for timed artificial insemination (TAI) in beef heifers (14-d CIDR-PGF(2α) program). The objective was to test a similar program in dairy cows by measuring first-service conception rates (FSCR), pregnancy rates after 2 AI, and time to pregnancy compared with a control (AI after observed estrus). Postpartum cows (Holstein, Jersey, or crossbred; n=1,363) from 4 grazing dairy farms were assigned to 1 of 2 programs: 14dCIDR_TAI [CIDR in for 14 d, CIDR out, PGF(2α) injection at 19 d after CIDR removal, GnRH injection 56 h later, and then TAI 16 h later; n=737] or control [AI after observed estrus; reproductive program with PGF(2α) (cycling cows) and CIDR (noncycling cows) to synchronize estrus with the start of the breeding season; n=626]. Body condition was scored (1 to 5; thin to fat) at the start of the trial. The interval from the start of the breeding period (final PGF(2α) injection of either program) to first AI was shorter for 14dCIDR_TAI compared with the control (3.0±0.2 vs. 5.3±0.2 d; mean ± SEM) but 14dCIDR_TAI cows had lesser FSCR than controls (48 vs. 61%). Farm affected FSCR (50, 51, 67, and 58% for farms 1 to 4). The BCS affected FSCR (50, 55, and 62% for BCS=2, 2.5, and 3, respectively). Cows that either calved the year before (carryover) or that calved early in the calving season had greater FSCR than cows that calved later in the calving season (55, 61, and 42%, respectively). The percentage of cows pregnant to AI (first and second inseminations within 31-d breeding season) was similar for 14dCIDR_TAI and control (64 vs. 70%) cows, but farm (64, 62, 80, and 69%) and time of calving (70, 76, and 56%: carryover, early, and late, respectively) affected the percentage. Survival analyses showed an initial advantage for 14dCIDR_TAI (more cows inseminated and more pregnancies achieved early in the breeding season) that was not maintained over time. Conclusions were that the 14dCIDR_TAI program achieved acceptable FSCR (48%) and overall AI pregnancy rates (64%), but did not surpass a control program that used AI after observed estrus (61 and 70%, respectively).

Impact of postpartum metritis on the regeneration of endometrial glands in dairy cows.

The postpartum uterus involutes to its pre-pregnant and fully functional state within approximately 60 d after calving. Uterine glands are essential for fertility but little is known about their regeneration postpartum. Likewise, the effect of uterine disease (metritis) on gland regeneration is unknown. We hypothesized that uterine glands would be regenerated early postpartum and that metritis would be associated with slower gland regeneration to affect their numbers later postpartum during the breeding period. Postpartum dairy cows were diagnosed as healthy (n = 17 and 9 for experiment [Exp.] 1 and 2) or metritis (n = 17 and 10 for Exp. 1 and 2, respectively) at 7 to 10 d postpartum. Cows were slaughtered at approximately 1 mo (Exp. 1) or approximately 80 or 165 d (Exp. 2) postpartum for the collection of the uterus. Uterine tissue was sectioned and the number of glandular cross-sections per unit area was counted and cross-sectional area measured. Cellular proliferation within the luminal epithelium (LE) and glandular epithelium (GE) was quantified by MKI67 (marker of cellular proliferation) immunohistochemistry. In early postpartum cows (Exp. 1), the greatest amount of MKI67 staining was found in the deep endometrium (cells closest to the myometrium). Cows with purulent material in the uterine lumen at d 30 slaughter (Exp. 1) had fewer endometrial glands per unit area in the deep and middle endometrium when compared with nonpurulent cows. The MKI67 staining was less in the deep endometrial GE and LE for purulent compared with nonpurulent cows. Estrus cyclicity was associated with a greater number of gland cross-sections in the deep and middle endometrium. Later postpartum (80 and 165 d; Exp. 2), there was greater glandular development compared with Exp. 1 and a tendency for a lesser number of gland cross-sections per unit area in diseased cows without an effect on MKI67 staining in the GE or LE. We conclude that uterine disease slows the development of uterine glands early postpartum (by 1 mo) through a mechanism that involves cellular proliferation within the GE. The impact of the early postpartum disease on glandular development later postpartum (Exp. 2) appeared to be less. Additional time, therefore, may allow recovery of the GE in later postpartum cows.

Short communication: presynchronization for timed artificial insemination in grazing dairy cows by using progesterone for 14 days with or without prostaglandin F2α at the time of progesterone withdrawal.

Progesterone-containing devices can be inserted intravaginally for 14 d to presynchronize the estrous cycle for timed artificial insemination (TAI) in beef heifers ("14-day CIDR-PG" or "Show-Me-Synch" program). The progesterone treatment is effective for presynchronization because cattle develop a persistent dominant follicle during treatment that ovulates within 3 d after progesterone removal. The subsequent estrous cycle can be effectively used for a TAI program. Some cattle will retain a functional corpus luteum (CL) for the entire 14-d treatment period and will not be synchronized effectively because the interval to ovulation depends on the lifespan of their existing CL. The objective was to test the effect of a luteolytic dose of PGF(2α) at progesterone removal for improving synchrony of estrus after treatment and increasing conception rate to a subsequent TAI in dairy cows. Postpartum cows (n = 1,021) from 2 grazing dairy herds were assigned to 1 of 2 presynchronization programs that used a controlled internal drug releasing (CIDR) device containing progesterone: 14dCIDR (CIDR in, 14 d, CIDR out; n = 523) or 14dCIDR+PGF(2α) (CIDR in, 14 d, CIDR out, and PGF(2α); n = 498). Cows were body condition scored (BCS; 1 to 5, thin to fat) and tail painted at CIDR removal. Paint score (PS) was recorded after CIDR removal [PS = 0 (all paint removed, indication of estrus), PS = 3 (paint partially removed), or PS = 5 (no paint removed; indication of no estrus)]. At 19 d after CIDR removal, all cows were treated with PGF(2α), 56 h later treated with GnRH, and then 16 h later were TAI. Treating cows with PGF(2α) at CIDR removal increased the percentage with PS = 0 within 5 d (58.1% vs. 68.9%; 14dCIDR vs. 14dCIDR+PGF(2α)). We found no effect of treatment, however, on conception rate at TAI (41.1% vs. 43.6%; respectively). The TAI conception rate increased with increasing BCS and was greater for cows that had PS = 0 within 5 d after CIDR removal. In summary, treating cows with PGF(2α) at CIDR removal increased the percentage of cows with all tail paint removed but did not increase percentage of pregnant cows after TAI.

Erratum to "Reproductive performance of early- and late-calving dairy cows artificially inseminated after ovulation synchronization and estrous resynchronization or artificially inseminated after observed estrus" (JDS Commun. 2:80-85).

[This corrects the article DOI: 10.3168/jdsc.2020-0035.].

Evaluation of the 7 & 7 Synch and 7-day CO-Synch + CIDR treatment regimens for control of the estrous cycle among beef cows prior to fixed-time artificial insemination with conventional or sex-sorted semen.

An experiment was conducted to compare the 7 & 7 Synch and 7-day CO-Synch + controlled internal drug release (CIDR®) treatment regimens before fixed-time artificial insemination (FTAI) of beef cows with conventional or sex-sorted semen. Cows (n = 1538) were blocked based on age and days postpartum (DPP) and randomly assigned to treatment regimen and semen type. Cows assigned to the 7-day CO-Synch + CIDR treatment regimen (n = 769) were administered gonadotropin-releasing hormone (GnRH) and an intravaginal progesterone-releasing insert (CIDR) on Day - 10, and administration of prostaglandin F2α (PG) coincident with CIDR removal on Day - 3. Cows assigned to 7 & 7 Synch (n = 769) were administered PG and a CIDR device on Day - 17, GnRH on Day - 10, and PG coincident with CIDR removal on Day - 3. Cows were administered GnRH coincident with FTAI, which was performed 66 h after CIDR removal with conventional (20 × 106 cells) or sex-sorted (4 × 106 cells) semen. Expression of estrus was affected by treatment regimen (P = 0.01) and by treatment regimen × DPP (P = 0.0004), as a result of imposing the 7 & 7 Synch regimen; therefore, a greater percentage of cows expressed estrus (82% compared with 64%), particularly among cows with greater DPP. Pregnancy percentages resulting from FTAI were less (P < 0.0001) when using sex-sorted semen but greater among cows treated with 7 & 7 Synch (conventional semen: 72%; sex-sorted semen: 52%) compared with 7-day CO-Synch + CIDR (conventional semen: 61%; sex-sorted semen: 44%).

Reproductive performance of early- and late-calving dairy cows artificially inseminated after ovulation synchronization and estrous resynchronization or artificially inseminated after observed estrus.

Within seasonal dairy systems, cows that calve late in the calving season are less likely to become pregnant and maintain a yearly calving interval. Very few studies have examined effective strategies for reproductive management of these late-calving cows. The objectives were to evaluate the reproductive performance of early- and late-calving dairy cows that were either inseminated after observed estrus (control) or enrolled in a timed AI and resynchronization protocol [progesterone (P4) Ovsynch Resynch)]. Early-calving cows calved during the first week of the calving season, whereas late-calving cows calved after 6 wk but were at least 10 d in milk at study commencement. Three dairy herds participated in the study with 391 cows total. Within each calving group, cows were randomly assigned to P4 Ovsynch Resynch or control (no treatment) in a 2 × 2 experimental design. Artificial insemination continued for 6 wk after mating start date (MSD) and was followed by 6 wk of natural service. The interval from MSD to AI was shorter (11.7 vs. 14.7 d) and the 3-wk pregnancy rate (49.5 vs. 21.2%) and the 6-wk pregnancy rate (60.8 vs. 42.4%) were greater in the early-calving compared with the late-calving control cows. By design, synchronized cows received timed AI on MSD and were not included in the statistical analysis of submission rate and interval from MSD to AI. The proportion of cows that received a second AI was not increased by the progesterone-based resynchronization strategy but was greater in early-calving compared with late-calving cows. The 12-wk pregnancy rate was greater (64.5 vs. 45.0%) in the early-calving synchronized cows compared with the late-calving synchronized cows. The interval from MSD to pregnancy was 6 and 12.6 d shorter for synchronized compared with control cows in the early- and late-calving groups, respectively. The results demonstrated reduced reproductive performance of late-calving cows compared with early-calving cows. Nonetheless, a major improvement to reproductive performance was achieved by targeting late-calving cows with a synchronization program, even when cows were only 20 to 50 DIM at first AI. Resynchronization of estrus with a progesterone device only, however, was not sufficient to increase the proportion of nonpregnant cows that received a second AI.

Growth of the conceptus from days 33 to 45 of pregnancy is similar for heifers and lactating cows and not associated with circulating glucose, insulin, IGF1 or progesterone concentrations.

Less than optimal circulating progesterone as well as glucose, insulin, and IGF1 concentrations in lactating cows may contribute to embryonic loss if placental development is dependent on concentrations of these hormones. There was comparison of fetal growth in lactating dairy cows with a contemporary group of nulliparous heifers (negative control for lactation). There was also assessment of the association between conceptus development and circulating hormone and metabolite concentrations and the response to an intravenous glucose administration. The embryo and amniotic vesicle were measured every 2-3 days during a 2-week period (days 33-45 of pregnancy). Blood samples were collected at the time of ultrasonography and analyzed for glucose, insulin, IGF1, and progesterone. There was also an intravenous glucose administration. Lactating cows had lesser blood concentrations of glucose, progesterone, and IGF1 during pregnancy. With respect to glucose administration, cows had lesser insulin and glucose than heifers at time 0 but heifers released three-fold more insulin than cows in response to glucose infusion. Cows had slightly larger fetuses when compared with heifers. When included in the model for embryo or amniotic vesicle length or area, concentrations of circulating glucose, insulin, IGF1, or progesterone did not explain a large amount of variation in values for these variables (P > 0.10). The conclusion was that cows differed from heifers for fetal growth but concentrations of circulating metabolites were not associated with size of the fetus. There was no evidence for an association between the hormonal or metabolic milieu and conceptus development.

Treatment with prostaglandin F2α and an intravaginal progesterone insert promotes follicular maturity in advance of gonadotropin-releasing hormone among postpartum beef cows.

An experiment was designed to evaluate treatments to promote ovarian follicular maturity in advance of administration of exogenous gonadotropin-releasing hormone (GnRH; 100 µg gonadorelin) for control of the bovine estrous cycle. We hypothesized prostaglandin F2α (PGF2α; 500 µg cloprostenol) followed by an intravaginal progesterone-releasing insert (CIDR; 1.38 g progesterone) would induce greater follicle size and serum estradiol at the time of GnRH administration. Postpartum cows (n = 194) in two locations were assigned to one of five treatments based on age, days postpartum, and body condition score. Cows in Treatment 1 were treated with the standard 7-d CO-Synch + CIDR protocol: administration of GnRH and CIDR insertion on Day -10, and administration of PGF2α and CIDR removal on Day -3. Treatments 2-5 were designed in a 2 × 2 factorial arrangement, with Treatment 1 included as an additional reference. On Day -17, cows in Treatments 2-5 received a CIDR insert, either with (Treatments 2 and 3) or without (Treatments 4 and 5) administration of PGF2α at CIDR insertion. On Day -10, all cows were administered GnRH, and CIDR inserts were either removed (Treatments 2 and 4) or remained in place until Day -3 (Treatments 3 and 5). Treatment with PGF2α and CIDR in advance of GnRH (Treatments 2 and 3) resulted in increased diameter of the largest ovarian follicle (P < 0.001) and increased serum concentrations of estradiol (P < 0.0005) on Day -10. In addition, variation among cows in CL status (no CL vs. a single CL vs. multiple CL) on Day -3 tended to be decreased (P = 0.08), with cows more likely to have a single CL rather than no CL or multiple CL. Lastly, the proportion of cows expressing estrus prior to fixed-time artificial insemination tended (P = 0.08) to be improved. Results support the hypothesis that administration of PGF2α and treatment with a CIDR for 7 days prior to GnRH promotes follicular maturity in advance of GnRH administration and may provide an approach by which to enhance response of postpartum beef cows to GnRH-based estrus synchronization programs.