Heterogeneous Fe-N-C Catalyst for Aerobic Dehydrogenation of Hydrazones to Diazo Compounds Used for Carbene Transfer.

Organic diazo compounds are versatile reagents in chemical synthesis and would benefit from improved synthetic accessibility, especially for larger scale applications. Here, we report a mild method for the synthesis of diazo compounds from hydrazones using a heterogeneous Fe-N-C catalyst, which has Fe ions dispersed within a graphitic nitrogen-doped carbon support. The reactions proceed readily at room temperature using O2 (1 atm) as the oxidant. Aryl diazoesters, ketones, and amides are accessible, in addition to less stable diaryl diazo compounds. Initial-rate data show that the Fe-N-C catalyst achieves faster rates than a heterogeneous Pt/C catalyst. The oxidative dehydrogenation of hydrazones may be performed in tandem with Rh-catalyzed enantioselective C-H insertion and cyclopropanation of alkenes, without requiring isolation of the diazo intermediate. This sequence is showcased by using a flow reactor for continuous synthesis of diazo compounds.

Evaluating differences in milk production, reproductive performance, and survival associated with vaginal discharge characteristics and fever in postpartum dairy cows.

The objective was to assess differences in productive and reproductive performance, and survival associated with vaginal discharge characteristics and fever in postpartum dairy cows located in Western and Southern states of the U.S.A. This retrospective cohort study included data from 3 experiments conducted in 9 dairies. Vaginal discharge was evaluated twice within 12 DIM and scored on a 5-point scale. The highest score observed for each cow was used for group assignment (VD group) as follows: VD 1 and 2 (VD 1/2; n = 1,174) = clear mucus/lochia with or without flecks of pus; VD 3 (n = 1,802) = mucopurulent with < 50% pus; VD 4 (n = 1,643) = mucopurulent with ≥50% of pus or non-fetid reddish/brownish mucous, n = 1,643; VD 5 = fetid, watery, and reddish/brownish, n = 1,800. All VD 5 cows received treatment according to each herd's protocol. Rectal temperature was assessed in a subset of VD 5 cows, and subsequently divided into Fever (rectal temperature ≥39.5°C; n = 334) and NoFever (n = 558) groups. A smaller proportion of cows with VD 5 (67.6%) resumed ovarian cyclicity compared with VD 1/2 (76.2%) and VD 4 (72.9%) cows; however, a similar proportion of VD5 and VD 3 (72.6%) cows resumed ovarian cyclicity. A smaller proportion of VD 5 (85.8%) cows received at least one artificial insemination (AI) compared with VD 1/2 (91.5%), VD 3 (91.0%), or VD 4 (91.6%) cows. Although we did not detect differences in pregnancy at first AI according to VD, fewer cows with VD 5 (64.4%) were pregnant at 300 DIM than cows with VD 1/2 (76.5%), VD 3 (76.2%), or VD 4 (74.7%). Hazard of pregnancy by 300 DIM was smaller for VD 5 compared with VD 1/2, VD 3, or VD 4 cows. A greater proportion of VD 5 cows were removed from the herd within 300 DIM compared with other VD groups. There was 760 kg lesser milk production within 300 DIM for VD 5 compared with VD 2, VD 3, and VD 4, whereas VD 2, VD 3, and VD 4 had similar milk production. We did not detect an association between fever at diagnosis of VD 5 and reproductive performance or milk production. A greater proportion of VD 5 cows without fever were removed from the herd by 300 DIM compared with VD 5 cows with fever. Differences in productive and reproductive performance, and removal of the herd were restricted to fetid, watery, and reddish/brownish vaginal discharge, which was independent of fever.

Chemical Kinetic Method for Active-Site Quantification in Fe-N-C Catalysts and Correlation with Molecular Probe and Spectroscopic Site-Counting Methods.

Mononuclear Fe ions ligated by nitrogen (FeNx) dispersed on nitrogen-doped carbon (Fe-N-C) serve as active centers for electrocatalytic O2 reduction and thermocatalytic aerobic oxidations. Despite their promise as replacements for precious metals in a variety of practical applications, such as fuel cells, the discovery of new Fe-N-C catalysts has relied primarily on empirical approaches. In this context, the development of quantitative structure-reactivity relationships and benchmarking of catalysts prepared by different synthetic routes and by different laboratories would be facilitated by the broader adoption of methods to quantify atomically dispersed FeNx active centers. In this study, we develop a kinetic probe reaction method that uses the aerobic oxidation of a model hydroquinone substrate to quantify the density of FeNx centers in Fe-N-C catalysts. The kinetic method is compared with low-temperature Mössbauer spectroscopy, CO pulse chemisorption, and electrochemical reductive stripping of NO derived from NO2- on a suite of Fe-N-C catalysts prepared by diverse routes and featuring either the exclusive presence of Fe as FeNx sites or the coexistence of aggregated Fe species in addition to FeNx. The FeNx site densities derived from the kinetic method correlate well with those obtained from CO pulse chemisorption and Mössbauer spectroscopy. The broad survey of Fe-N-C materials also reveals the presence of outliers and challenges associated with each site quantification approach. The kinetic method developed here does not require pretreatments that may alter active-site distributions or specialized equipment beyond reaction vessels and standard analytical instrumentation.

Transcriptome changes associated with elongation of bovine conceptuses I: Differentially expressed transcripts in the conceptus on day 17 after insemination.

The objective was to characterize transcriptome changes associated with elongation in bovine conceptuses during preimplantation stages. Nonlactating Holstein cows were euthanized 17 d after artificial insemination (AI) and the uterine horn ipsilateral to the CL was flushed with saline solution. Recovered conceptuses were classified as small (1.2 to 6.9 cm; n = 9), medium (10.5 to 16.0 cm; n = 9), or large (18.0 to 26.4 cm; n = 10). Total mRNA was extracted and subjected to transcriptome analyses using the Affymetrix Gene Chip Bovine array. Data were normalized using the GCRMA method and analyzed by robust regression using the Linear Models for Microarray library within Bioconductor in R. Transcripts with P ≤ 0.05 after adjustment for false discovery rate and fold change ≥1.5 were considered differentially expressed. Functional analyses were conducted using the Ingenuity Pathway Analysis platform. Comparisons between large versus small (LvsS), large versus medium (LvsM), and medium versus small (MvsS) conceptuses yielded a total of 634, 240, and 63 differentially expressed transcripts, respectively. Top canonical pathways of known involvement with embryo growth that were upregulated in large conceptuses included actin cytoskeleton (LvsS), integrin signaling (LvsS and LvsM), ephrin receptor (LvsS), mesenchymal transition by growth factor (LvsM), and regulation of calpain protease (LvsS). Transcripts involved with lipid metabolism pathways (LXR/RXR, FXR/RXR, hepatic fibrosis) were associated with the LvsS and LvsM, and some transcripts such as APOC2, APOH, APOM, RARA, RBP4, and PPARGC1A, were involved in these pathways. An overall network summary associated biological downstream effects of invasion of cells, proliferation of embryonic cells, and inhibition of organismal death in the LvsS. In conclusion, differently expressed transcripts in the LvsS comparison were associated with the cell growth, adhesion, and organismal development, although part of these findings could be attributed to differences in circulatory concentrations of progesterone of the cows that bore large and small conceptuses. The large and medium conceptuses developed under similar concentrations of progesterone and presented 240 differently expressed transcripts, associated with cell differentiation, metabolite regulation, and other biological processes.

Quinone-mediated hydrogen anode for non-aqueous reductive electrosynthesis.

Electrochemical synthesis can provide more sustainable routes to industrial chemicals1-3. Electrosynthetic oxidations may often be performed 'reagent-free', generating hydrogen (H2) derived from the substrate as the sole by-product at the counter electrode. Electrosynthetic reductions, however, require an external source of electrons. Sacrificial metal anodes are commonly used for small-scale applications4, but more sustainable options are needed at larger scale. Anodic water oxidation is an especially appealing option1,5,6, but many reductions require anhydrous, air-free reaction conditions. In such cases, H2 represents an ideal alternative, motivating the growing interest in the electrochemical hydrogen oxidation reaction (HOR) under non-aqueous conditions7-12. Here we report a mediated H2 anode that achieves indirect electrochemical oxidation of H2 by pairing thermal catalytic hydrogenation of an anthraquinone mediator with electrochemical oxidation of the anthrahydroquinone. This quinone-mediated H2 anode is used to support nickel-catalysed cross-electrophile coupling (XEC), a reaction class gaining widespread adoption in the pharmaceutical industry13-15. Initial validation of this method in small-scale batch reactions is followed by adaptation to a recirculating flow reactor that enables hectogram-scale synthesis of a pharmaceutical intermediate. The mediated H2 anode technology disclosed here offers a general strategy to support H2-driven electrosynthetic reductions.

Transcriptome changes associated with elongation of bovine conceptuses II: Differentially expressed transcripts in the endometrium on day 17 after insemination.

The objective was to characterize endometrial transcriptome on d 17 of gestation in dairy cows according to conceptus length. Nonlactating Holstein cows (n = 48) were slaughtered 17 d after AI and the uterine horn ipsilateral to the corpus luteum (CL) was flushed with saline solution. Recovered conceptuses were classified as small (1.2-6.9 cm; n = 9), medium (10.5-16.0 cm; n = 9), or large (18.0-26.4 cm; n = 10). Samples of intercaruncular endometrium dissected from the caudal, intermediate, and cranial portions of the uterine horn ipsilateral to the pregnancy were pooled for analyses. Total mRNA was extracted from endometrial tissue and subjected to transcriptome analyses using the Affymetrix Gene Chip Bovine array. Data were normalized using the GCRMA method and analyzed by robust regression using the Linear Models for Microarray library within Bioconductor in R. Transcripts with P ≤ 0.05 after adjustment for false discovery rate and fold change ≥1.5 were considered differentially expressed. Functional analyses were conducted using the Ingenuity Pathway Analysis platform. Comparisons between endometria of cows carrying large versus small (LvsS), large versus medium (LvsM), and medium versus small (MvsS) conceptuses yielded a total of 235, 21, and 94 differentially expressed transcripts, respectively. Top canonical pathways included the antigen presentation pathway and Th1/Th2 activation pathways, both for LvsS and MvsS. Interferon-α and -γ were identified as activated upstream regulators, primarily based on differently expressed transcripts such as IDO1, ISG20, WARS, LGALS9, IFI44, and PSMB9 (LvsS and MvsS). For LvsS, regulator analyses revealed predicted activation of FOXO1, IFN, NFACTC2, IL-12, IL-6, and IL-18, whereas it depicted inhibition of IL10RA and ZBTB1. Changes in these regulators were associated with a downstream activation of leukocytes, as well as quantity and expansion of T lymphocytes. Canonical pathways associated with the comparison LvsM included cell cycle G2/M DNA damage checkpoint regulation, cell cycle control of chromosomal replication. Moreover, tretinoin was predicted, as activated in upstream analysis for the same comparison. In conclusion, most of the differently expressed transcripts in the endometrium on d 17 of gestation were identified between cows carrying small conceptuses compared with counterparts carrying medium and large conceptuses and were involved with pathways associated with modulation of the immune response.

Combined effect of purulent vaginal discharge and anovulation on pregnancy status in a large multi-state population of Holstein cows.

The objective of this observational prospective cohort study was to evaluate the combined effect of purulent vaginal discharge (PVD) and anovulation (ANOV) on the reproductive performance of a large multi-state population of Holstein cows. Data were prospectively collected from 11,729 cows in 16 herds located in 4 regions in the United States [Northeast (4 herds), Midwest (6), Southeast (1), and Southwest (5)]. Cows were enrolled at calving and monitored weekly for disease occurrence, reproductive events, and survival. Prevalence of PVD was evaluated at 28 ± 3 d in milk and defined by the presence of mucopurulent to fetid vaginal discharge. Resumption of ovarian cyclicity was determined via transrectal ultrasonography at 40 ± 3 and 54 ± 3 d postpartum. Pregnancy diagnosis was performed by ultrasonography on d 32 ± 3 after artificial insemination (AI) and reconfirmed at d 60 ± 3 of gestation. Pregnancy loss (PL) was defined as a cow diagnosed pregnant at 32 ± 3 but nonpregnant at 60 ± 3 d after AI. The association of PVD and ANOV with pregnancy traits was analyzed using 4 PVD-cyclicity categories that considered the following combinations: NPVD-CYC = absence of PVD and cycling; PVD-CYC = presence of PVD and cycling; NPVD-ANOV = absence of PVD and anovular; and PVD-ANOV = presence of PVD and anovular. Multiple logistic regression and Cox proportional regression were used for the analysis of potential associations between PVD and cyclicity categories and pregnancy at first AI (PAI1), days from calving to pregnancy, and PL at first AI. The odds (95% confidence intervals) of pregnancy increased from cows in the PVD-ANOV category (reference category) to cows in NPVD-ANOV [2.09 (1.62-2.50)], PVD-CYC [2.52 (2.02-3.14)], and NPVD-CYC [3.46 (2.84-4.23)]. Similarly, days from calving to pregnancy were less for NPVD-CYC, followed by PVD-CYC, NPVD-ANOV, and PVD-ANOV (121.4, 137.2, 137.3, and 157.4 d, respectively). On the contrary, no clear association was identified between groups and PL. The results indicate that both PVD and ANOV had a negative impact on PAI1 and days from calving to pregnancy. The results indicated a variable magnitude in the negative impact on the reproductive traits analyzed when both conditions were combined.

Heterogeneous M-N-C Catalysts for Aerobic Oxidation Reactions: Lessons from Oxygen Reduction Electrocatalysts.

Nonprecious metal heterogeneous catalysts composed of first-row transition metals incorporated into nitrogen-doped carbon matrices (M-N-Cs) have been studied for decades as leading alternatives to Pt for the electrocatalytic O2 reduction reaction (ORR). More recently, similar M-N-C catalysts have been shown to catalyze the aerobic oxidation of organic molecules. This Focus Review highlights mechanistic similarities and distinctions between these two reaction classes and then surveys the aerobic oxidation reactions catalyzed by M-N-Cs. As the active-site structures and kinetic properties of M-N-C aerobic oxidation catalysts have not been extensively studied, the array of tools and methods used to characterize ORR catalysts are presented with the goal of supporting further advances in the field of aerobic oxidation.

Effects of maternal choline supplementation on performance and immunity of progeny from birth to weaning.

The objectives were to investigate whether supplementation with rumen-protected choline (RPC) during late pregnancy in Holstein cows affects offspring immunity and growth, and whether effects are utero-placental, colostrum dependent, or both. A total of 105 multiparous Holstein cows were assigned randomly to a prepartum diet (1.54 Mcal of NEL/kg of DM, and 15.8% CP) without (control) or with added RPC (12.9 g/d of choline ion). Calves (n = 111) were blocked by sex and assigned randomly to colostrum from control cows or colostrum from RPC cows, resulting in 4 treatments in a 2 × 2 factorial arrangement: (1) calves born and fed colostrum from non-supplemented dams (NN; n = 33); (2) calves from non-supplemented dams and fed colostrum from RPC-fed cows (NC; n = 25); (3) calves from RPC-supplemented dams and colostrum from non-supplemented cows (CN; n = 28); and (4) calves from RPC-supplemented dams and colostrum from RPC-fed cows (CC; n = 25). Growth, intakes, and immunity of females were evaluated up to 56 d of age. Growth and intake of male calves was evaluated up to 35 d of age, and physiological and immune responses to intravenous LPS challenge were evaluated from 21 to 35 d of age. Effects of prenatal and colostrum treatments and interactions between treatments were analyzed using mixed models. Calves fed colostrum from RPC-supplemented dams had a 17.4% increase in apparent efficiency of absorption of IgG compared with calves fed colostrum from control dams (27.4 vs. 23.3%). Incidence of fever in the first 21 d of age tended to be less in females born from RPC-supplemented dams compared with females born from control dams (31 vs. 58%). Prenatal RPC females had increased hematocrit and concentrations of red blood cells, leukocytes, neutrophils, and lymphocytes in blood compared with prenatal females born from control dams. Compared with prenatal control females, prenatal RPC females had greater intake of milk replacer (704 vs. 748 ± 9.9 g/d) and starter (45.4 vs. 60.2 ± 5.9 g/d) during the first 21 d of age. In male calves, mean intake of DM was greater (1,074 vs. 976 ± 45 g/d) after the LPS challenge (0 to 8 d) by calves born from dams fed RPC compared with males born from control dams. Calves born from RPC-fed dams also had lower mean rectal temperature (39.0 vs. 39.2°C) and mean respiration rate (35.6 vs. 39.3 breaths/min) compared with males born from control dams. Moreover, serum concentrations of metabolites (i.e., ß-hydroxybutyric acid, fatty acids, and glucose), cytokines (i.e., tumor necrosis factor-α) and acute phase proteins (i.e., serum amyloid A) were consistent with less-severe inflammatory response to LPS in males born from dams fed RPC compared with control. Source of colostrum and interaction between prenatal and colostrum treatments had minimal effects on calf responses to LPS. Overall, maternal RPC supplementation during late gestation suggests a positive effect on immunity, in that colostrum from RPC-fed dams increased efficiency of IgG absorption and maternal supplementation with RPC during late gestation, regardless of colostrum source, attenuated responses to LPS.

Molecular Catalyst Synthesis Strategies to Prepare Atomically Dispersed Fe-N-C Heterogeneous Catalysts.

We report a strategy to integrate atomically dispersed iron within a heterogeneous nitrogen-doped carbon (N-C) support, inspired by routes for metalation of molecular macrocyclic iron complexes. The N-C support, derived from pyrolysis of a ZIF-8 metal-organic framework, is metalated via solution-phase reaction with FeCl2 and tributyl amine, as a Brønsted base, at 150 °C. Fe active sites are characterized by 57Fe Mössbauer spectroscopy and aberration-corrected scanning transmission electron microscopy. The site density can be increased by selective removal of Zn2+ ions from the N-C support prior to metalation, resembling the transmetalation strategy commonly employed for the preparation of molecular Fe-macrocycles. The utility of this approach is validated by the higher catalytic rates (per total Fe) of these materials relative to established Fe-N-C catalysts, benchmarked using an aerobic oxidation reaction.

Heat stress during the transition period is associated with impaired production, reproduction, and survival in dairy cows.

Objectives of this retrospective cohort study were to determine the association of heat stress (HS) exposure during the periparturient period with production, health, reproduction, and survival during the first 90 d postpartum in dairy cows. A total of 5,722 Holstein cows (2,324 nulliparous and 3,397 parous) were categorized into environmental condition groups based on average temperature-humidity index (THI) exposure as thermoneutral (TN) or HS during the prepartum (PRE) and early postpartum (POST) periods into TN-TN (THI: PRE = 57.9 and POST = 63.7), TN-HS (THI: PRE = 68.2 and POST = 72.0), HS-TN (THI: PRE = 71.4 and POST = 65.6), and HS-HS (THI PRE = 72.2 and POST = 72.7). Nulliparous and parous cows were analyzed separately. In nulliparous cows, exposure to HS during the PRE, POST, or both PRE and POST periods was associated with a 1.7 kg/cow per day reduction in milk yield compared with TN-TN. Postpartum HS was associated with increases of 4.4 percentage points in incidence of retained placenta, 18.1 percentage points in incidence of metritis, and 2.0 percentage points in incidence of mastitis, but a reduction of 5.3 percentage points in pregnancy at first AI, and an increase of 4.5 percentage points in pregnancy loss compared with POST TN. Exposure to HS during PRE and POST periods was associated with increased removal from the herd. In parous cows, exposure to HS during PRE, POST, or both PRE and POST was associated with a 2.4-kg/cow per day reduction in milk yield when compared with TN-TN. There was an interaction between HS exposure PRE and POST and incidence of retained placenta, because POST HS was associated with an increase in incidence of 5.8 percentage points within PRE HS cows, whereas no difference was found within PRE TN cows. Postpartum HS was associated with an increase of 6.3 percentage points in incidence of metritis and tended to be associated with an increase of 1.9 percentage points in mastitis incidence compared with POST TN. There was an interaction between HS exposure PRE and POST and pregnancy per AI because POST HS was associated with reduced pregnancy incidence by 10.6 percentage points within PRE TN cows, whereas no difference was found within PRE HS cows. Removal from the herd increased in cows exposed to HS during the PRE or POST or PRE and POST. These data suggest that POST HS is associated with performance losses to a greater extent than prepartum HS and that nulliparous and parous cows are prompt to losses associated with exposure to HS during the transition period. The results corroborate findings from manipulative experiments that showed improved milk production and reproduction in cows under HS provided with heat abatement but adds insights to the associations between HS and health.

Chemical and Electrochemical O2 Reduction on Earth-Abundant M-N-C Catalysts and Implications for Mediated Electrolysis.

M-N-C catalysts, incorporating non-precious-metal ions (e.g. M = Fe, Co) within a nitrogen-doped carbon support, have been the focus of broad interest for electrochemical O2 reduction and aerobic oxidation reactions. The present study explores the mechanistic relationship between the O2 reduction mechanism under electrochemical and chemical conditions. Chemical O2 reduction is investigated via the aerobic oxidation of a hydroquinone, in which the O-H bonds supply the protons and electrons needed for O2 reduction to water. Mechanistic studies have been conducted to elucidate whether the M-N-C catalyst couples two independent half-reactions (IHR), similar to electrode-mediated processes, or mediates a direct inner-sphere reaction (ISR) between O2 and the organic molecule. Kinetic data support the latter ISR pathway. This conclusion is reinforced by rate/potential correlations that reveal significantly different Tafel slopes, implicating different mechanisms for chemical and electrochemical O2 reduction.

Effects of progesterone concentrations and follicular wave during growth of the ovulatory follicle on conceptus and endometrial transcriptome in dairy cows.

Objectives were to evaluate the effects of follicular wave and progesterone concentration on growth of the ovulatory follicle, conceptus elongation, uterine IFN-τ concentration, and transcriptome of conceptus and endometrium of pregnant cows on d 17 of gestation. Nonlactating nonpregnant Holstein cows were assigned randomly to one of 3 treatments: ovulation of a first-wave follicle (FW, n = 15); ovulation of a first-wave follicle and progesterone supplementation (FWP4, n = 12); and ovulation of a second-wave follicle (SW, n = 19). Ovulation of a first- or second-wave follicle was achieved by initiating the Ovsynch protocol (d -9 GnRH, d -2 and -1 PGF2α, d 0 GnRH and artificial insemination, d 0.7 artificial insemination) on d 0 or 6 of a presynchronized estrous cycle, respectively. Cows in FWP4 received 3 intravaginal inserts containing progesterone at 12, 24, and 48 h after the first GnRH injection that were removed on d -2. Cows were killed on d 17 for collection of the reproductive tract. Transcriptome was evaluated by microarray using the Affymetrix Bovine Array. Orthogonal contrasts were built to assess the effects of progesterone concentration during follicle growth (FW vs. FWP4 + SW) and follicular wave (FWP4 vs. SW). Progesterone concentrations (LSM ± SEM) from d -9 to -2 were greater for SW, followed by FWP4 and FW (5.38 ± 0.24, 4.26 ± 0.28, and 1.17 ± 0.27 ng/mL). Diameter of the ovulatory follicle (FW = 19.6 ± 0.6; FWP4 = 15.6 ± 0.6; SW = 15.2 ± 0.5 mm) and concentrations of estradiol from d -2 to 1 (FW = 4.05 ± 0.33; FWP4 = 2.73 ± 0.35; SW = 2.48 ± 0.30 pg/mL) were greater for FW compared with FWP4 and SW. Progesterone concentrations from d 3 to 16 were greater for FW compared with FWP4 and SW. A total of 28 singleton conceptuses were collected (FW, n = 8; FWP4, n = 8; SW, n = 12) and only intact conceptuses were included in the analyses of length (FW, n = 8; FWP4, n = 6; SW, n = 12). Although conceptuses were longer for FW compared with FWP4 and SW (FW = 16.6 ± 2.3; FWP4 = 9.8 ± 2.2; SW = 9.6 ± 2.0 cm), treatment did not affect the amount of IFN-τ in uterine flushing. Transcriptome of conceptuses and endometrium of pregnant cows was not extensively affected by follicular wave (8 and 1 differentially expressed transcripts) or concentration of progesterone during follicle growth (0 and 3 differentially expressed transcripts), showing that these factors did not affect conceptuses and endometrium transcriptome in pregnancies that are maintained to d 17.

Effect of body condition change and health status during early lactation on performance and survival of Holstein cows.

Body condition score (BCS) and disease records are commonly available in dairy operations. However, the effect of BCS changes (ΔBCS) considering specific health profiles has not been investigated extensively. The objective of this study was to assess the effects of different levels of ΔBCS on fertility, milk yield, and survival of Holstein cows diagnosed with reproductive disorders (REP; dystocia, twins, retained fetal membranes, metritis, and clinical endometritis), other health disorders (OTH; subclinical ketosis, left displaced abomasum, lameness, clinical mastitis, and respiratory disease), or with no disease events (HLT) within 40 days in milk (DIM). Data included lactation information from 11,733 cows calving between November 2012 and October 2014 in 16 herds across 4 geographical regions in the United States (Northeast, Midwest, Southwest, Southeast). Cows were evaluated for BCS at 5 ± 3 DIM (BCS5) and at 40 ± 3 DIM (BCS40) and the difference between BCS40 and BCS5 was classified as excessive loss of BCS (EL; ΔBCS ≤-0.75), moderate loss (ML; ΔBCS = -0.5 to -0.25), no change (NC; ΔBCS = 0), or gain of BCS (GN; ΔBCS ≥0.25). Multivariable logistic regression was used for assessing potential associations between the outcomes of interest and ΔBCS and health. The effect of the interaction term ΔBCS by health group was not statistically significant for any of the study outcomes. The odds of resumption of ovarian cyclicity (ROC), in GN, NC, and ML cows were 1.94 (95% CI: 1.57-2.40), 1.59 (1.28-1.97), and 1.27 (1.10-1.47) times greater than the odds of ROC in EL cows, respectively. The odds of pregnancy at 150 DIM (P150) in GN cows were 1.61 (1.20-2.17) times greater than the odds of P150 in EL cows. Cows with REP or OTH disorders had smaller odds of ROC compared with HLT cows [REP: OR = 0.65 (0.56-0.76) and OTH: OR = 0.79 (0.68-0.92)]. For pregnancy outcomes, REP cows had smaller odds of pregnancy at the first artificial insemination compared with HLT cows [0.70 (0.58-0.84)]. Similarly, REP cows had smaller odds of being diagnosed pregnant by 150 and 305 DIM compared with HLT cows [P150: 0.73 (0.59-0.87), P305: 0.58 (0.49-0.69)]. Overall, average daily milk within the first 90 DIM was greater in EL (39.5 ± 1.13 kg/d) and ML (38.9 ± 1.11 kg/d) cows than in NC (37.8 ± 1.12 kg/d) and GN (36.2 ± 1.12 kg/d) cows. On the other hand, average daily milk within the first 90 DIM was lower in REP (37.0 ± 1.11 kg/d) cows compared with OTH (38.7 ± 1.12 kg/d) and HLT cows (38.6 ± 1.11 kg/d). The magnitude of ΔBCS and the health status of early lactation cows should be considered when assessing subsequent cow performance and survival.

Peripheral leucocyte molecular indicators of inflammation and oxidative stress are altered in dairy cows with embryonic loss.

Objective of experiment was to determine whether oxidative stress (OS) and inflammation altered embryonic loss in dairy cows. Blood samples were collected at days 0, 16, 32 and 60 after timed (AI) from 200 Holstein cows to determine embryonic loss based on interferon-stimulated gene-15 (ISG15) mRNA expression (day 16) and ultrasound at day 32 and day 60. Leucocyte expressions of mRNA TLR2, TLR4, TNF-α, IL1B, IL10, STAT3 (inflammation), PTGS2, PTGES (prostaglandin synthesis), and PLA2G4A and ALOX5AP (eicosanoid metabolism) at days 0 and 16 were determined. Plasma redox status for antioxidant enzymatic activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), total antioxidant capacity (TAC), and concentrations of malondialdehyde (MDA) were determined at days 0, 16, 32 and 60. All antioxidant-redox responses were beneficially significant in pregnant cows diagnosed pregnant at day16 and sustained pregnancy to day 60 compared to non-pregnant cows at day16 or pregnant at day16 and lost embryos by days 32 or 60. The leucocyte mRNA expressions of TLR2, TLR4, STAT 3, IL1B, PTGS2, PLA2G4A and ALOX5AP were greater and PTGES was lower at day16 in pregnant cows that lost embryos early (P < 0.05). In conclusion peripheral leucocyte molecular indicators of inflammation and plasma indicators of OS were altered in pregnant cows undergoing embryonic losses compared to cows with a sustained pregnancy.

Effects of maternal level of dietary cation-anion difference fed to prepartum nulliparous cows on offspring acid-base balance, metabolism, and growth.

The objectives were to determine the effects of dietary cation-anion difference (DCAD) fed to pregnant cows during the last 22 d of gestation on offspring acid-base balance, metabolism, growth, and health preweaning. A total of 132 nulliparous Holstein cows were enrolled at 250 (248 to 253) d of gestation in a randomized block design. Cows were blocked by genomic merit of energy-corrected milk yield and assigned randomly to diets varying in DCAD: +200 (P200, n = 43), -50 (N50, n = 45), or -150 (N150, n = 44) mEq/kg of dry matter (DM). Newborn calves (15 males and 28 females in P200, 22 males and 23 females in N50, and 18 males and 26 females in N150) were followed for the first 7 or 56 d of age if males or females, respectively. Measures of acid-base balance and concentrations of minerals in blood were measured in all calves on d 0 before colostrum feeding, and on d 1, 3, and 7. Each calf was fed 3.78 L of colostrum from the respective treatment, and apparent efficiency of IgG absorption was determined. All calves were weighed at birth, and females were weighed again at 21, 42, and 56 d of age. Concentrations in serum of total calcium (tCa), total magnesium (tMg), and total phosphorus (tP) were measured up to 56 d of age; intakes of milk and starter grain DM were measured daily from 21 to 56 d of age; and incidence of disease was recorded for the first 56 d of age in females. Treatment did not affect acid-base balance measured in all calves. Calves were born with metabolic and respiratory acidosis, which reversed by 1 d of age. In the first 24 h after birth, blood pH increased from 7.215 to 7.421 and bicarbonate from 26.2 to 31.7 mM, whereas partial pressure of CO2 decreased from 64.1 to 48.7 mm of Hg in all treatments. Maternal DCAD did not affect colostrum IgG content fed to calves (P200 = 95.0 vs. N50 = 91.0 vs. N150 = 97.1 ± 4.1 g/L) or apparent efficiency of IgG absorption (P200 = 33.1 vs. N50 = 33.1 vs. N150 = 34.2 ± 1.9%). Males were born heavier than females, but maternal DCAD did not affect birth weight of all calves (P200 = 37.7 vs. N50 = 37.3 vs. N150 = 37.8 ± 0.7 kg) or daily weight gain in females in the first 56 d of life (P200 = 0.80 vs. N50 = 0.81 vs. N150 = 0.77 ± 0.03 kg/d). Treatment did not affect intake of milk (P200 = 1.11 vs. N50 = 1.04 vs. N150 = 1.19 ± 0.06 kg/d) or starter grain DM (P200 = 0.27 vs. N50 = 0.27 vs. N150 = 0.21 ± 0.06 kg/d), or measures of feed efficiency. Treatment did not affect concentrations of minerals in serum, morbidity, or age at morbidity. Manipulating the DCAD of pregnant nulliparous dams during late gestation did not affect offspring performance in the first 2 mo of age.

Days in the prepartum group are associated with subsequent performance in Holstein cows.

Our objective was to evaluate the association between days in the prepartum group (DPG) with performance and survival in Holstein cows. Data from 18,657 Holstein cow-lactations (6,993 nulliparous and 9,390 parous prepartum) were collected. Cows with a gestation length shorter than 256 d (n = 267) or longer than 296 d (n = 131) and cows that spent 0 DPG (n = 238) were removed, resulting in 18,021 cow-lactations. Data were collected for the first 300 d postpartum, and responses included milk yield, incidence of diseases by 90 d postpartum, reproduction, and survival. Days in the prepartum group were analyzed as a continuous variable, and regression coefficients were used to estimate the responses when cows spent 7, 28, or 42 DPG, representing cows with a short, moderate, or an extended time in the prepartum group, respectively. An interaction between DPG as a quadratic covariate and parity-diet was observed for milk yield by 300 d postpartum. Means were 9,331; 9,665; and 9,261 kg for 7, 28, or 42 DPG, respectively, in nulliparous cows, and 9,886; 10,939; and 10,117 kg for 7, 28, or 42 DPG, respectively, in parous cows. Also, the interaction between DPG and parity-diet affected retained placenta, metritis, mastitis, and morbidity. Morbidity affected 49.5, 52.9, and 59.5% of nulliparous and 49.7, 26.5, and 47.4% of parous cows that spent 7, 28, or 42 DPG, respectively. A linear association between DPG and pregnancy at first artificial insemination was observed with estimates of 37.0, 32.6, and 29.8% for 7, 28, and 42 DPG, respectively. On the other hand, a quadratic association was observed between DPG and the proportion of pregnant cows at 300 d postpartum, and estimates for 7, 28, and 42 DPG were, respectively, 71.7, 73.5, and 58.8%. A quadratic relationship was also observed for DPG and removal from the herd by 300 d postpartum, and estimates were 25.2, 22.9, and 34.4% for 7, 28, or 42 DPG, respectively. Associations between DPG with production, health, reproduction, and survival were detected, and they varied with parity-diet group. For several responses evaluated, a quadratic association was observed, which suggested that there was an optimal period for cows to spend in the prepartum group, and reduced or extended number of days were detrimental to performance.

The economic cost of metritis in dairy herds.

The objective of this study was to estimate the cost of metritis in dairy herds. Data from 11,733 dairy cows from 16 different farms located in 4 different regions of the United States were compiled for up to 305 d in milk, and 11,581 cows (2,907 with and 8,674 without metritis) were used for this study. Metritis was defined as fetid, watery, red-brownish vaginal discharge that occurs ≤21 d in milk. Continuous outcomes such as 305-d milk production, milk sales ($/cow), cow sales ($/cow), metritis treatment costs ($/cow), replacement costs ($/cow), reproduction costs ($/cow), feeding costs ($/cow), and gross profit per cow ($/cow) were analyzed using mixed effect models using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Gross profit was also compared using the Kruskal-Wallis test. Dichotomous outcomes such as pregnant and culling by 305 d in milk were analyzed using the GLIMMIX procedure of SAS. Time to pregnancy and culling were analyzed using the PHREG procedure of SAS. Models included the fixed effects of metritis, parity, and the interaction between metritis and parity, and farm as the random effect. Variables were considered significant when P ≤ 0.05. Metritis cost was calculated by subtracting the gross profit of cows with metritis from the gross profit of cows without metritis. A stochastic analysis was performed with 10,000 iterations using the observed results from each group. Milk yield and proportion of cows pregnant were lesser for cows with metritis than for cows without metritis, whereas the proportion of cows leaving the herd was greater for cows with metritis than for cows without metritis. Milk sales, feeding costs, residual cow value, and gross profit were lesser for cows with metritis than for cows without metritis. Cow sales and replacement costs were greater for cows with metritis than for cows without metritis. The mean cost of metritis from the study herds was $511 and the median was $398. The stochastic analysis showed that the mean cost of a case of metritis was $513, with 95% of the scenarios ranging from $240 to $884, and that milk price, treatment cost, replacement cost, and feed cost explained 59%, 19%, 12%, and 7%, respectively, of the total variation in cash flow differences. In conclusion, metritis caused large economic losses to dairy herds by decreasing milk production, reproduction, and survival in the herd.

Duration and degree of diet-induced metabolic acidosis prepartum alter tissue responses to insulin in dairy cows.

The objective of this experiment was to determine the effects of altering the dietary cation-anion difference (DCAD) fed for the last 21 or 42 d of gestation on glucose metabolism and tissue insulin responsiveness. Ninety parous Holstein cows at 232 d of gestation were assigned randomly to dietary treatments with 2 levels of DCAD (-70 or -180 mEq/kg) fed for 2 durations (short: the last 21 d of gestation; long: the last 42 d of gestation). For the short treatments, a diet with +110 mEq/kg was fed from 232 to 254 d of gestation. Intravenous glucose tolerance tests (IVGTT) were performed at either 250 or 270 d of gestation by infusing 0.25 g of dextrose/kg of body weight within 1 min. The following day, cows underwent an insulin challenge (IC) and received 0.1 IU of insulin/kg of body weight intravenously. Blood was sampled at min -15, -5, and 0 to establish a baseline and from 5 to 180 min relative to infusions; plasma concentrations of glucose, insulin, and fatty acids were determined, and the respective areas under the curves (AUC) were calculated. Liver was sampled after the IVGTT, and adipose tissue was sampled after the IVGTT and IC for quantification of mRNA expression and protein abundance. Reducing the DCAD altered acid-base balance compatible with a compensated metabolic acidosis. At 250 d, reducing the DCAD increased the AUC for glucose and reduced that of insulin following the IVGTT, whereas during the IC, clearance rate decreased and time to half-life of insulin increased with reducing DCAD, resulting in a tendency to a larger AUC for fatty acids. At 270 d, quantitative insulin sensitivity check index and the revised quantitative insulin sensitivity check index were smaller in cows fed the acidogenic diets for the last 42 d of gestation compared with the last 21 d of gestation, thereby suggesting reduced insulin sensitivity. In addition, cows fed for the long duration tended to have greater AUC for glucose but smaller AUC for insulin following an IVGTT than those fed for the short duration, thereby suggesting reduced insulin release and glucose disposal. Treatments did not affect hepatic mRNA expression of G6PC, PCK1, PCK2, and PC or adipose tissue mRNA expression of ATGL, ACC, B2AR, HSL, and PLIN1. On the other hand, for proteins, reducing the DCAD linearly reduced abundance of rabbit anti-mouse protein kinase B (AKT) and tended to reduce rabbit anti-human phosphorylated (Ser-9) glycogen synthase kinase-3 ß (pGSK) and the pGSK:rabbit anti-human glycogen synthase kinase-3 ß (GSK) ratio in hepatic tissue, whereas a linear increase in rabbit anti-human hormone-sensitive lipase (HSL) and rabbit anti-mouse phosphorylated (Ser-660) hormone-sensitive lipase (pHSL) in adipose tissue was observed after the IVGTT at 250 d. Moreover, reducing the DCAD resulted in a linear reduction of AKT and tended to reduce rabbit anti-human acetyl-CoA carboxylase (ACC) but increased pHSL linearly in adipose tissue after an IC at 250 d. Cows fed acidogenic diets for a short duration tended to have less pHSL in adipose tissue than those fed for a long duration after an IVGTT at 270 d. Associations were observed between blood pH and mRNA and protein abundance in hepatic and adipose tissues. Diet-induced metabolic acidosis altered insulin release and insulin signaling, resulting in a shift in adipose tissue metabolism that would favor lipolysis over lipogenesis.

Effects of injectable calcitriol on mineral metabolism and postpartum health and performance in dairy cows.

Objectives were to determine the effects of an injectable formulation of calcitriol on Ca concentration, risk of clinical diseases, and performance in dairy cows. Cows were blocked by lactation number (1 vs. >1) and calving sequence and, within block, assigned randomly within 6 h of calving to receive subcutaneously vehicle only (CON, n = 450) or 200 (CAL200, n = 450) or 300 µg of 1α,25-dihydroxyvitamin D3 (CAL300, n = 450). Cows were fed the same acidogenic diet prepartum. Blood was sampled before treatment administration and again during the first 11 d postpartum and analyzed for concentrations of ionized Ca (iCa), total Ca (tCa), Mg (tMg), and P (tP), ß-hydroxybutyrate, carboxylated osteocalcin (cOC), and undercarboxylated osteocalcin (uOC). Cows were evaluated for diseases in the first 60 d postpartum. Reproduction and survival were monitored for the first 300 d postpartum. Calcitriol increased concentration of blood iCa (CON = 1.12 vs. CAL200 = 1.23 vs. CAL300 = 1.27 mM), plasma tCa (CON = 2.29 vs. CAL200 = 2.44 vs. CAL300 = 2.46 mM), and plasma tP (CON = 1.72 vs. CAL200 = 2.21 vs. CAL300 = 2.28 mM), and differences were observed during the first 5 d postpartum for iCa and tCa, and the first 7 d postpartum for tP. Concentrations of tMg were lower in calcitriol-treated cows than in CON cows (CON = 0.81 vs. CAL200 = 0.78 vs. CAL300 = 0.75 mM), and differences were observed during the first 5 d postpartum. Calcitriol increased plasma concentrations of cOC (CON = 14.5 vs. CAL200 = 23.0 vs. CAL300 = 19.8 ng/mL) and uOC (CON = 1.6 vs. CAL200 = 3.4 vs. CAL300 = 2.6 ng/mL). Prevalence of subclinical hypocalcemia was less in calcitriol-treated cows (CON = 19.0 vs. CAL200 = 4.7 vs. CAL300 = 9.3%); however, benefits on health were only observed in overconditioned cows (n = 270/1,350). Calcitriol reduced incidence of retained placenta (CON = 14.3 vs. CAL200 = 5.1 vs. CAL300 = 5.9%), puerperal metritis (CON = 12.7 vs. CAL200 = 6.1 vs. CAL300 = 2.5%), and morbidity (CON = 72.1 vs. CAL200 = 57.4 vs. CAL300 = 56.9%) in cows with BCS greater than 3.50, but no benefit on health was observed in cows with BCS equal to or less than 3.50 at parturition. Milk yield did not differ among treatments. Pregnancy at first AI did not differ, but pregnancy rate after the first AI was slower for calcitriol-treated cows because of reduced insemination rate and pregnancy per AI. We found that CAL200 reduced death but increased culling in cows without calving problems. Collectively, results indicate that treatment with calcitriol at parturition was effective in improving concentrations of iCa, tCa, and tP, which reduced the risk of hypocalcemia. Pregnancy rate was reduced by calcitriol treatment, and benefits on health performance were limited to overconditioned cows. Thus, treatment of all cows is not supported, and proper identification of cohorts of cows that benefit from postpartum interventions that increase blood calcitriol or calcium is needed.