Short communication: Reproduction outcomes in dairy heifers following a 14-d progesterone insert presynchronization protocol.

The objectives were to evaluate pregnancy per artificial insemination (AI), days to first AI, and proportion pregnant within 7 d of AI eligibility in dairy heifers subjected to presynchronization compared with dairy heifers not presynchronized. Thirty days before AI eligibility, Holstein heifers were assigned randomly to 1 of 3 groups: 14-d controlled internal drug release (CIDR; containing progesterone) presynchronization, PGF2α presynchronization, or control (no presynchronization). Heifers in the 14-d CIDR presynchronization treatment (n = 119) received a CIDR on d -30, which was removed on d -16, followed by an injection of PGF2α upon entry to the breeding program (d 0). Heifers in the PGF2α presynchronization treatment (n = 118) received an injection of PGF2α on d -11 and d 0. Control heifers (n = 121) were not presynchronized and received an injection of PGF2α on d 0. All heifers received tail paint on d 0 to facilitate once-daily detection of estrus (based on paint removal). Heifers detected in estrus received AI with conventional semen on the same morning as detected estrus. Generalized linear mixed models were used to assess mean treatment differences. Following PGF2α treatment on d 0, more heifers were detected in estrus in the first 7 d after eligibility in the 14-d CIDR group (95.8%) compared with the PGF2α (74.6%) and control (66.9%) groups. Days to first AI differed between treatments (14-d CIDR = 3.6 d vs. PGF2α = 5.0 d vs. control = 6.8 d). Pregnancy per AI for first AI within 7 d of eligibility was 71.9% (14-d CIDR), 58.0% (PGF2α), and 61.7% (control), and differed between 14-d CIDR and PGF2α heifers. Presynchronization with a 14-d CIDR increased the proportion of heifers pregnant in the first 7 d of eligibility (14-d CIDR = 68.9% vs. PGF2α = 43.2% vs. control = 41.3%). Projected days on feed (d 0 to projected calving date) were 295 (14-d CIDR), 302 (PGF2α), and 305 (control), and were different between the 14-d CIDR and control heifers. The potential economic benefit to the producer was $15.85 per heifer presynchronized with a 14-d CIDR protocol compared with the control group. Treatment of dairy heifers with a 14-d CIDR effectively presynchronized estrus, resulting in a greater proportion detected in estrus, reduced days to first AI, and an increased proportion of heifers pregnant within the first 7 d after breeding eligibility compared with heifers presynchronized with a single PGF2α injection and control heifers.

Pregnancy outcomes are not improved by administering gonadotropin-releasing hormone at initiation of a 5-day CIDR-Cosynch resynchronization protocol for lactating dairy cows.

Using a 5-d controlled internal drug-release (CIDR)-Cosynch resynchronization protocol, the objective of this study was to determine the effect of the initial GnRH injection on pregnancy per artificial insemination (P/AI) to the second artificial insemination in lactating Holstein dairy cows. On 37 ± 3 d (mean ± standard deviation) after the first artificial insemination, and upon nonpregnancy diagnosis (d 0 of the experiment), lactating cows eligible for a second artificial insemination (n = 429) were enrolled in a 5-d CIDR-Cosynch protocol. On d 0, all cows received a CIDR insert and were assigned randomly to receive the initial GnRH injection (GnRH; n = 226) of the protocol or no-GnRH (n = 203). Blood samples were collected from a sub-group of cows (n = 184) on d 0 and analyzed for progesterone (P4) concentration. On d 5, CIDR inserts were removed, and all cows received 1 injection of PGF2α. On d 6 and 7, cows were observed once daily by employees for tail-chalk removal, and cows detected in estrus on d 6 or 7 received artificial insemination that day (EDAI), and did not receive the final GnRH injection. The remaining cows not detected in estrus by d 8 received GnRH and timed artificial insemination (TAI). Pregnancy status was confirmed by transrectal palpation of uterine contents at 37 ± 3 d (mean ± standard deviation) after the second artificial insemination. Eliminating the initial GnRH injection had no effect on P/AI compared with cows receiving GnRH (27 vs. 21%), respectively. Similarly, method of insemination (EDAI vs. TAI) and its interaction with treatment had no effect on P/AI. Primiparous cows had greater P/AI than multiparous cows (31 vs. 21%). Mean P4 concentrations (n = 184) at the initiation of the protocol did not differ between treatments (4.51 ± 0.35 ng/mL no-GnRH vs. 3.96 ± 0.34 ng/mL of GnRH). When P4 concentrations were categorized as high (≥1 ng/mL) or low (<1 ng/mL), P/AI tended to be greater for high P4 concentrations (n = 136) compared with low (n = 48) P4 concentrations (26 vs. 16%, respectively). No differences were observed in the proportion of cows with high or low P4 between treatments. Collectively, these results provide evidence that eliminating the initial GnRH in a 5-d CIDR-Cosynch resynchronization protocol for lactating dairy cows did not reduce P/AI in this study.

Human milk oligosaccharides promote the growth of staphylococci.

Human milk oligosaccharides (HMO), which constitute a major component of human milk, promote the growth of particular bacterial species in the infant's gastrointestinal tract. We hypothesized that HMO also interact with the bacterial communities present in human milk. To test this hypothesis, two experiments were conducted. First, milk samples were collected from healthy women (n = 16); culture-independent analysis of the bacterial communities was performed, HMO content was analyzed, and the relation between these factors was investigated. A positive correlation was observed between the relative abundance of Staphylococcus and total HMO content (r = 0.66). In a follow-up study, we conducted a series of in vitro growth curve experiments utilizing Staphylococcus aureus or Staphylococcus epidermidis and HMO isolated from human milk. HMO exhibited stimulatory effects on bacterial growth under various nutritional conditions. Analysis of culture supernatants from these experiments revealed that HMO did not measurably disappear from the culture medium, indicating that the growth-enhancing effects were not a result of bacterial metabolism of the HMO. Instead, stimulation of growth caused greater utilization of amino acids in minimal medium. Collectively, the data provide evidence that HMO may promote the growth of Staphylococcus species in the lactating mammary gland.

Short communication: characterization of early postpartum estrous behavior and ovulation in lactating dairy cows using radiotelemetry.

The objective of this study was to describe early postpartum estrous behavior and ovulation in lactating dairy cows using radiotelemetry. Cows (n=50) were continuously monitored for behavioral estrus with a radiotelemetric system, HeatWatch II (CowChips LLC, Manalapan, NJ), from d 14 to approximately d 49 postpartum. Blood collection for analysis of progesterone and ovarian ultrasonography were performed once weekly starting on d 14. First ovulation was associated with behavioral estrus in 5 cows and occurred at 28.2±10.8 d (mean±SD; range 17 to 40 d). The average duration of estrus was 6.0±4.9 h (range 3 to 12.2 h), and the mean number of standing events was 18.4±8.9 (range 4 to 26). Based on progesterone concentrations of ≥1 ng/mL, estimated first postpartum ovulation occurred at 25.1±10.4 d (range 10 to 49 d) for 38 animals without evidence of behavioral estrus. The interval to estimated first ovulation without behavioral estrus was not different from the interval to first ovulation associated with behavioral estrus. Level of milk production and body condition score loss did not affect the interval to estimated first ovulation without estrus or first ovulation associated with estrus. Six animals did not show evidence of ovulation based on progesterone concentration, whereas 1 cow showed evidence of estrous behavior on the day before removal from the study. The majority of first postpartum ovulations (38/43; 88.4%) were not associated with behavioral estrus.

Pregnancy per artificial insemination during summer in lactating dairy cows after treatment with aspirin.

Embryonic loss in cattle may be related to a hormonal imbalance resulting in alterations in timing of prostaglandin F2α (PGF2α) secretion around the time of maternal recognition of pregnancy. The objective of this study was to examine effects of aspirin (a PGF2α inhibitor) on pregnancy per AI (P/AI), and progesterone (P4), and pregnancy specific protein B (PSPB) concentrations in lactating dairy cows inseminated more than once after parturition. Fourteen days after second or subsequent AI (Day 0 = Day of AI), 556 cows were assigned randomly to aspirin (187.2 g total; n = 277) or control (n = 279) groups. Aspirin was administered orally on Day 14 and 15, and control cows were subjected to sham bolus administration. On Day 25, blood samples were collected from a subset of cows (n = 194) to quantify P4 and PSPB, whereas pregnancy was determined in all cows at 35-42 days post-AI. Maximum daily ambient temperature ranged from 38-41 °C during the experiment. Mean parity, days in milk, and times bred before treatment (TBRD) did not differ between groups. There were no differences in P/AI between treatments (aspirin 21.6 % compared with control 27.5 %). Neither treatment, parity, TBRD, or any two-way interactions with treatment affected concentrations of P4. Moreover, there were no effects (P > 0.50) of treatment, or treatment by TBRD interaction on serum PSPB concentrations. A tendency (P = 0.07) occurred for multiparous cows to have greater serum PSPB concentrations compared with primiparous cows. Mean serum PSPB concentrations tended (P = 0.07) to be greater for second or third TBRD compared to fourth and greater TBRD. These results provide evidence that aspirin administered during periods of heat stress after the second and subsequent AI post-partum during the summer months does not improve P/AI or alter P4 and PSPB in lactating dairy cows.

Fatty acids and monoacylglycerols inhibit growth of Staphylococcus aureus.

Staphylococcus aureus causes a variety of human infections including toxic shock syndrome, osteomyelitis, and mastitis. Mastitis is a common disease in the dairy cow, and S. aureus has been found to be a major infectious organism causing mastitis. The objectives of this research were to determine which FA and esterified forms of FA were inhibitory to growth of S. aureus bacteria. FA as well as their mono-, di-, and triacylglycerol forms were tested for their ability to inhibit a human toxic shock syndrome clinical isolate (MN8) and two S. aureus clinical bovine mastitis isolates (305 and Novel). The seven most potent inhibitors across all strains tested by minimum inhibitory concentration analysis included lauric acid, glycerol monolaurate, capric acid, myristic acid, linoleic acid, cis-9, trans-11 conjugated linoleic acid, and trans-10, cis-12 conjugated linoleic acid. Some of these lipids were chosen for 48-h growth curve analysis with a bovine mastitis S. aureus isolate (Novel) at doses of 0, 20, 50, and 100 microg/mL except myristic acid, which was tested at 0, 50, 100, and 200 microg/mL. The saturated FA (lauric, capric, myristic) and glycerol monolaurate behaved similarly and reduced overall growth. In contrast, the polyunsaturated FA (linoleic and cis-9, trans-11 conjugated linoleic acid) delayed the time to initiation of exponential growth in a dose-dependent fashion. The results suggest that lipids may be important in the control of S. aureus during an infection.

Estrogen receptor-directed radiotoxicity with Auger electrons: specificity and mean lethal dose.

To assess the feasibility of using estrogen receptor-directed therapy with Auger electron-emitting ligands for therapy of estrogen receptor (ER)-containing cancers, we synthesized and evaluated the radiotoxicity of several 123I-labeled estrogens to specifically kill ER+ cells in culture. Auger electrons have been previously shown to be of short range, generally less than the dimensions of a cell, so that to use them therapeutically a mechanism is needed to deliver the Auger electron-emitting nuclide to the vicinity of the DNA. Since it is now well established that the estrogen receptor, when bound to estrogen, forms a high affinity association with distinct estrogen response elements in the DNA, we wished to test the hypothesis that a short exposure of cells to a 123I-labeled estrogen would be specifically radiotoxic to ER+ cells, and that the decays per cell needed for cell killing would be compatible with reasonable levels of receptor occupancy. Using the halodestannylation reaction with tributyl tin precursors of several estrogens and commercially available iodine-123, we prepared the iodoestrogens, E-17 alpha(-)[123I]-iodo-11 beta-methoxyestradiol and 2(-)[123I]iodo-1,1-bis(4-hydroxyphenyl)--2-phenylethylene, at high specific activities, in several cases at essentially the specific activity of 123I itself, 240,000 Ci/mmol. When various concentrations of either of the 123I-labeled estrogens were incubated for 1 h with a subline of ER+ Chinese hamster ovary cells and the washed cells plated for survival assays, a dose-dependent, unlabeled estradiol-inhibitable reduction in survival was observed. In contrast, Chinese hamster ovary cells not expressing estrogen receptor showed little sensitivity to the radiotoxicity of the 123I-labeled estrogens. Calculations based on the assayed residence time of the iodoestrogens in the cells indicate that several hundred decays per cell are sufficient to kill cells.

A meta-analysis on the relationship between intake of nutrients and body weight with milk volume and milk protein yield in dairy cows.

Previously observed strong relationships between dry matter (DM) intake and milk yield in dairy cows were the basis for this meta-analysis aimed to determine the influence of intake of specific dietary nutrients on milk yield and milk protein yield in Holstein dairy cows. Diets (563) from feeding trials published in the Journal of Dairy Science were evaluated for nutrient composition using 2 diet evaluation programs. Intake of nutrients was estimated based on DM intake and program-derived diet composition. Data were analyzed with and without the effect of stage of lactation. Models based on intake of nutrients improved prediction of milk yield and milk protein yield compared with DM intake alone. Intake of net energy of lactation was the dominant variable in milk yield prediction models derived from both diet evaluation models. Milk protein yield models also improved prediction over the DM intake model. These models were dominated by ruminally undegradable protein intake and included a number of energy-related intake variables. In most models, incorporating stage of lactation improved the model fit.

Short communication: Conception rates following detection of estrus and timed AI in dairy cows synchronized using GnRH and PGF2alpha.

The objective of this study was to compare conception rates of cows exhibiting spontaneous estrus and receiving artificial insemination (AI) before completion of a timed AI protocol with cows that did not display estrus spontaneously, but were inseminated after 1 of 3 GnRH-PGF2alpha protocols. Cows (n = 432) in 2 herds were administered GnRH on d -7 and were tail-chalked daily. Cows detected in estrus before d 0 were inseminated immediately. Cows not detected in estrus by d 0 were administered PGF2alpha and were tail-chalked daily until 48 h after PGF2alpha. Cows detected in estrus from d -7 to 48 h after PGF2alpha were inseminated and designated as treatment A (n = 46). Cows not detected in estrus and not inseminated by 48 h after PGF2alpha were assigned randomly to receive either GnRH 48 h after PGF2alpha and timed AI 16 h later (treatment B; n = 132), or GnRH and timed AI 64 h after PGF2alpha (treatment C; n = 127), or timed AI 64 h after PGF2alpha (treatment D; n = 127). Pregnancy was diagnosed 38 to 45 d after AI by palpation per rectum of uterine contents. Nearly 11% of all cattle exhibited spontaneous estrus and received immediate AI. Herd did not influence the percentage of cows detected in estrus and inseminated. Conception rates did not differ among treatments. Conception rates differed between herds, but no interaction of herd x treatment was detected. No differences were detected between herds for days in milk, milk production, AI service number, or parity.

Estimation of repeatability of blood constituents in gestating beef cattle on protein- and energy-restricted diets.

Repeatabilities of blood constituents were calculated for 104 Angus heifers on two separate experiments fed adequate, protein-deficient, energy-deficient, or both protein- and energy-deficient diets. Four statistical methodologies were compared including analysis of variance, principal component (structural) analysis based on the sample covariance and sample correlation matrix, and maximum likelihood. Of 12 blood constituents tested only seven were considered sufficiently important to be included in the analysis. These blood constituents included blood urea nitrogen (BUN), creatinine (Creat), alkaline phosphatase (Alk Phos), total protein (T Prot), total bilirubin (T Bil), cholesterol (Chol) and Iron (Fe). If the standard linear model assumptions were met for heifers on the adequate diet, the estimators appeared to be quite similar for both years except when the correlation coefficient was relatively small. If the assumption of homogeneity of the variance-covariance matrix (compound symmetry) was relaxed, the structural analysis method based on the sample correlation matrix appeared preferable. However, when combining all diets, the maximum likelihood methodology was preferred. Among the specific blood constituents, Alk Phos had the highest repeatability, not only for the heifers on the adequate diet, but also for heifers on other treatments in both years. Repeatabilities for T Prot appeared to be the most consistent over all rations in both years. Repeatability estimates for Fe were high and relative rankings were consistent for both years, while repeatabilities for the other variables were either low and(or) inconsistent.

Effect of clinical mastitis and other diseases on reproductive performance of Holstein cows.

The objective of this study was to evaluate the effect of clinical mastitis and (or) other diseases on reproductive performance in lactating Holstein cows. Cows (n=967) from a commercial dairy farm were divided into four groups retrospectively: cows with clinical mastitis and other diseases (MD, n=54), clinical mastitis only (M, n=154), other diseases only (D, n=187), and cows with no record of clinical mastitis or other diseases (H, n=572). Days in milk at first service (DIMFS), services per conception (S/C), days not pregnant (DNP), the rate at which animals became pregnant over time and the proportion of cows that remained non-pregnant during 224 days of lactation were evaluated. Groups MD and M had greater (P<0.05) DNP compared with H (155+/-15 and 140+/-5 vs. 88+/-2, respectively). Moreover, MD and M had greater (P<0.05) S/C compared with H (3.0+/-0.4 and 2.1+/-0.1 vs. 1.6+/-0.1, respectively). The rate at which animals became pregnant over time was less (P<0.05) for MD and M and tended (P=0.1) to be less for D when compared with H. In addition, proportion of cows that remained non-pregnant by 224 days of lactation was greater (P<0.05) in MD, M, and D compared with H. Cows with mastitis were also divided into three groups according to the day of occurrence of the first case of clinical mastitis: (1) clinical mastitis occurred before 56 days postpartum (MP1); (2) clinical mastitis occurred between 56 and 105 days after parturition (MP2); and (3) clinical mastitis occurred after 105 days postpartum (MP3) Regardless of the time of occurrence, DNP was greater (P<0.05) for cows with mastitis compared with H. Time of mastitis occurrence affected S/C in that cows in MP2 and MP3 had a greater S/C compared with H cows (P<0.05). Reproductive efficiency was decreased by the presence of clinical mastitis alone because a greater proportion of cows with mastitis remained non-pregnant over time. Moreover, a greater proportion of cows with mastitis or diseases remained non-pregnant by 224 postpartum. Furthermore, the negative effects on reproduction were exacerbated when cows experienced both clinical mastitis and other diseases.

A meta-analysis examining the relationship among dietary factors, dry matter intake, and milk and milk protein yield in dairy cows.

This meta-analysis was undertaken to determine the impact of dietary components on dry matter intake (DMI), milk yield (MY), and milk protein yield (MPY) in Holstein dairy cows. Diets (n=846) from 256 feeding trials published in Volumes 73 through 83 of the Journal of Dairy Science were evaluated for nutrient composition using 2 diet evaluation models: CPM Dairy (a computer program based on the principles of the Cornell Net Carbohydrate and Protein System) and NRC (2001). Data were analyzed with and without the effect of stage of lactation as a dummy variable (<100 d in milk or > or =100 d in milk). A mixed model regression analysis was used to completely investigate the potential relationships among composition variables and DMI, MY, and MPY. Protein and carbohydrate fractions were the main components within the DMI models, and DMI played a dominant role in estimating MY and MPY. Inclusion of stage of lactation substantially improved the MY models but did not affect model fits or residual structure for DMI and MPY.

Effect of simultaneous thawing of multiple 0.5-mL straws of semen and sequence of insemination on conception rate in dairy cattle.

The objectives of this study were to determine 1) the effect of simultaneous thawing of multiple 0.5-mL straws of semen and sequence of insemination (first, second, third, or fourth) on conception rates in dairy cattle, 2) whether the conception rates achieved following AI by professional AI (PAI) technicians and herdsman-inseminators (HI) differed, and 3) the effect of elapsed time from initiation of thawing straws of semen to seminal deposition on conception rates in dairy cattle. Four dairies with PAI and four with HI participated in the study. Initial data recorded included beginning thaw time, cow identification number, and time of seminal deposition. Herd records were retrieved following pregnancy diagnosis. Conception rates of dairy cows (n = 1025) were not affected by sequence of insemination (first, second, third, or fourth). Conception rates for herds using PAI were 40, 47, 41, and 50%. Conception rates for herds using HI were 24, 20, 33, and 30%. Average conception rates of dairy cows differed between PAI and HI (45 vs. 27%, respectively). The difference in mean conception rate achieved by PAI and HI was not attributable to milk production, parity, service number or stage of lactation. The elapsed time from initial thaw to completion of fourth AI was shorter for PAI than for HI, 7.6 vs. 10.9 min, respectively. Although the average conception rate differed between PAI and HI, elapsed time from initial thaw to completion of fourth AI and sequence of insemination (first, second, third, or fourth) had no effect on conception rate within inseminator group.

Effect of dose of calcium salts of conjugated linoleic acid (CLA) on percentage and fatty acid content of milk fat in midlactation holstein cows.

Increasing conjugated linoleic acid (CLA) content of milk fat from lactating dairy cattle has become a research interest due to the possible health benefits afforded humans consuming CLA. Dietary supplementation of CLA to lactating dairy cows is one potential method by which CLA content of milk and dairy products may be enhanced. Feeding CLA in calcium salt form could potentially deliver CLA to the lower digestive tract through prevention of biohydrogenation by rumen microbes. Milk fat depression (MFD) occurs when cows receive CLA-60, a commercially available CLA source containing numerous CLA isomers, abomasally. Our objectives were to determine the quantity of CLA as calcium salts required to elicit maximal MFD and to evaluate the effects of CLA supplementation on fatty acid composition of milk fat. Five Holstein cows at approximately 93 DIM were utilized in a 5 x 5 balanced Latin square crossover design. Periods were 14-d in length with a 5-d treatment phase and 9-d rest phase. Treatments were 5-d supplementation of 0, 12.5, 25, 50, and 100 g of CLA-60 in calcium salt form. Milk samples were collected on d 5 of CLA supplementation and analyzed for composition and fatty acid profile. Regression analysis of milk fat data suggested that MFD was not maximized over the dose levels investigated, despite delivery of 34.5 g of trans-10, cis-12 CLA in the 100-g dose of CLA. Supplementation with 50 and 100 g of CLA per day resulted in a reduction of milk fat percent of 29 and 34%, respectively. Trend analysis indicated a linear decrease in the milk fat content of caprylic, capric, and lauric acids as the dose of CLA increased. Milk fat content of cis-9, trans-11, and trans-10, cis-12 CLA increased at an increasing rate as dose increased.