Short communication: Short-term effect of 3-nitrooxypropanol on feed dry matter intake in lactating dairy cows.

The objective of this study was to investigate the effect of 3-nitrooxypropanol (3-NOP), an enteric methane inhibitor under investigation, on short-term dry matter intake (DMI) in lactating dairy cows. Following a 1-wk adaptation period, 12 multiparous Holstein cows were fed a basal total mixed ration (TMR) containing increasing levels of 3-NOP during 5 consecutive, 6-d periods. The experiment was conducted in a tiestall barn. Feed bins were split in half by a solid divider, and cows simultaneously received the basal TMR supplemented with the following: (1) a placebo without 3-NOP or (2) 3-NOP included in the TMR at 30, 60, 90, or 120 mg/kg of feed dry matter (experimental periods 2, 3, 4, and 5, respectively). Cows received the control diet (basal TMR plus placebo premix) during experimental period 1. A premix containing ground corn grain, soybean oil, and dry molasses was used to incorporate 3-NOP in the ration. Cows were fed twice daily as follows: 60% of the daily feed allowance at 0800 h and 40% at 1800 h. Feed offered and refused was recorded at each feeding. During the morning feedings, each cow was offered either control or 3-NOP-treated TMR at 150% of her average intake during the previous 3 d. After collection of the evening refusals, cows received only the basal TMR without the premix until the next morning feeding. The test period for the short-term DMI data collection was defined from morning feeding to afternoon refusals collection during each day of each experimental period. Location (left or right) of the control and 3-NOP diets within a feed bin was switched every day during each period to avoid feed location bias. Dry matter intake of TMR during the test period was quadratically increased by 3-NOP compared with the control. Inclusion of 3-NOP at 120 mg/kg of feed dry matter resulted in decreased 10-h DMI compared with the lower 3-NOP doses, but was similar to the control. There was no effect of feed location (left or right) within feed bin on DMI. Data from this short-term study suggests that 3-NOP does not have a negative effect on DMI in lactating dairy cows.

Dose-response effect of 3-nitrooxypropanol on enteric methane emissions in dairy cows.

This experiment was designed to test the effect of inclusion rate of 3-nitrooxypropanol (3-NOP), a methane inhibitor, on enteric methane emissions in dairy cows. The study was conducted with 49 multiparous Holstein cows in a randomized complete block design in 2 phases; phase 1 was with 28 cows, and phase 2 with 21 cows. Cows were fed a basal total mixed ration ad libitum and were blocked based on days in milk, milk yield, and enteric methane emissions during a 14-d covariate period. Treatments were control (no 3-NOP) and 40, 60, 80, 100, 150, and 200 mg of 3-NOP/kg of feed dry matter. Following a 14-d adaptation period, enteric gaseous emissions (methane, carbon dioxide, and hydrogen) were measured using the GreenFeed system (C-Lock Inc., Rapid City, SD) over a 3-d period. Compared with the control, inclusion rate of 3-NOP quadratically decreased daily enteric methane emissions from 22 to 40%. Maximum mitigation effect was achieved with the 3 highest 3-NOP doses (with no statistical difference among 100, 150, and 200 mg/kg). The decrease in methane emission yield and emission intensity ranged from 16 to 36% and from 25 to 45%, respectively. Emissions of hydrogen quadratically increased 6- to 10-fold, compared with the control; the maximum increase was with 150 mg/kg 3-NOP. Treatment did not affect daily emissions of carbon dioxide, but a linear increase in carbon dioxide emission yield was observed with increasing 3-NOP doses. Dry matter intake and milk yield of the cows was not affected by 3-NOP. Milk fat concentration and yield were increased by 3-NOP due to increased concentration of de novo synthetized short-chain fatty acids in milk. Inclusion of 3-NOP also tended to increase milk urea nitrogen but had no other effects on milk components. In this short-term experiment, 3-NOP decreased enteric methane emissions without affecting dry matter intake or milk yield and increased milk fat in dairy cows. Maximum mitigation effect was achieved at 100 to 200 mg/kg of feed dry matter.

Implementation of a Brazilian Cardioprotective Nutritional (BALANCE) Program for improvement on quality of diet and secondary prevention of cardiovascular events: A randomized, multicenter trial

Background Complex percutaneous coronary intervention (PCI) is associated with higher ischemic risk, which can be mitigated by long-term dual antiplatelet therapy (DAPT). However, concomitant high bleeding risk (HBR) may be present, making it unclear whether short- or long-term DAPT should be prioritized. Objectives This study investigated the effects of ischemic (by PCI complexity) and bleeding (by PRECISE-DAPT [PRE dicting bleeding Complications in patients undergoing stent Implantation and Sub sequent Dual Anti Platelet Therapy] score) risks on clinical outcomes and on the impact of DAPT duration after coronary stenting. Methods Complex PCI was defined as ≥3 stents implanted and/or ≥3 lesions treated, bifurcation stenting and/or stent length >60 mm, and/or chronic total occlusion revascularization. Ischemic and bleeding outcomes in high (≥25) or non-high (<25) PRECISE-DAPT strata were evaluated based on randomly allocated duration of DAPT. Results Among 14,963 patients from 8 randomized trials, 3,118 underwent complex PCI and experienced a higher rate of ischemic, but not bleeding, events. Long-term DAPT in non-HBR patients reduced ischemic events in both complex (absolute risk difference: −3.86%; 95% confidence interval: −7.71 to +0.06) and noncomplex PCI strata (absolute risk difference: −1.14%; 95% confidence interval: −2.26 to −0.02), but not among HBR patients, regardless of complex PCI features. The bleeding risk according to the Thrombolysis In Myocardial Infarction scale was increased by long-term DAPT only in HBR patients, regardless of PCI complexity. Conclusions Patients who underwent complex PCI had a higher risk of ischemic events, but benefitted from long-term DAPT only if HBR features were not present. These data suggested that when concordant, bleeding, more than ischemic risk, should inform decision-making on the duration of DAPT. (AU)

Production effects of feeding extruded soybean meal to early-lactation dairy cows.

The objective of this experiment was to evaluate productive and reproductive effects of replacing solvent-extracted soybean meal (SSBM) with extruded soybean meal (ESBM) in a total mixed ration for early-lactation dairy cows. Thirty-four Holstein cows (12 primiparous and 22 multiparous) were used in a randomized complete block design experiment with 17 cows per treatment. Feeding was ad libitum for 5 to 10% refusals. A fresh-cow diet was fed the first 21 d in milk followed by a lactation diet from 22 to 60 d in milk. Milk and dry matter intake data were collected throughout the experiment, and samples were collected for blood chemistry and amino acid profile, nutrient digestibility, nitrogen utilization, and enteric methane emission using the GreenFeed system (C-Lock Inc., Rapid City, SD). Dry matter intake, milk yield, and feed efficiency were not different between SSBM and ESBM. Energy-corrected milk yield and efficiency were also not different between diets. Diet had no effect on milk composition, except that milk true protein yield was decreased by ESBM. Enteric methane emission, yield, and intensity were not different between SSBM and ESBM. Because of its greater fat content, ESBM triggered expected changes in milk fatty acid (FA) profile: decreased sum of C16, saturated, and odd- and branched-chain FA and increased sum of preformed FA, polyunsaturated, and trans FA. The ESBM diet increased or tended to increase some essential amino acids in plasma. In this study, ESBM did not affect dry matter intake and did not improve lactational performance or onset of ovarian function in early-lactation dairy cows, and it decreased milk protein yield, possibly due to greater unsaturated FA intake compared with SSBM.

Effect of dietary crude protein degradability and corn processing on lactation performance and milk protein composition and stability.

The present study aimed to evaluate the effect of crude protein degradability and corn processing on lactation performance, milk protein composition, milk ethanol stability (MES), heat coagulation time (HCT) at 140°C, and the efficiency of N utilization for dairy cows. Twenty Holstein cows with an average of 162 ± 70 d in milk, 666 ± 7 kg of body weight, and 36 ± 7.8 kg/d of milk yield (MY) were distributed in a Latin square design with 5 contemporaneous balanced squares, 4 periods of 21 d, and 4 treatments (factorial arrangement 2 × 2). Treatment factor 1 was corn processing [ground (GC) or steam-flaked corn (SFC)] and factor 2 was crude protein (CP) degradability (high = 10.7% rumen-degradable protein and 5.1% rumen-undegradable protein; low = 9.5% rumen-degradable protein and 6.3% rumen-undegradable protein; dry matter basis). A significant interaction was observed between CP degradability and corn processing on dry matter intake (DMI). When cows were fed GC with low CP degradability, DMI increased by 1.24 kg/d compared with cows fed GC with high CP degradability; however, CP degradability did not change DMI when cows were fed SFC. Similar interactions were observed for MY, HCT, and lactose content. When cows were fed GC diets, high CP degradability reduced MY by 2.3 kg/d, as well as HCT and lactose content, compared with low CP degradability. However, no effect of CP degradability was observed on those variables when cows were fed SFC diets. The SFC diets increased dry matter and starch total-tract digestibility and reduced ß-casein (CN) content (% total milk protein) compared with GC diets. Cows fed low-CP degradability diets had higher glycosylated κ-CN content (% total κ-CN) and MES, as well as milk protein content, 3.5% fat-corrected milk, and efficiency of N for milk production, than cows fed high-CP degradability diets. Therefore, GC and high-CP degradability diets reduced milk production and protein stability. Overall, low CP degradability increased the efficiency of dietary N utilization and MES, probably due to changes in casein micelle composition, as CP degradability or corn processing did not change the milk concentration of ionic calcium. The GC diets increased ß-CN content, which could contribute to reducing HTC when cows were fed GC and high-CP degradability diets.

Acceptance of flavored concentrate premixes by young ruminants following a short-term exposure.

Three cafeteria feeding design experiments were conducted to test whether young ruminants have flavor preferences. Experiment 1 was with 11 Dorset × Suffolk weaned lambs of both sexes, aged 5 mo and averaging 47.5 (standard deviation = 5.8) kg of body weight. The lambs were offered a choice of 5 flavored concentrate premixes (FCP) and an unflavored control for 5 min 4 times over 10 d. The FCP were prepared by mixing 200 to 300 g/t (as-is basis) of synthetic flavors (vanilla, milky, spicy/fenugreek, red summer fruits, and molasses) into a basal diet. The unflavored control and the milky flavor were consumed in greater amounts than all other flavors at 83.9 and 65.8 g/test, respectively. The consumption rate of FCP (g/min) was similar among treatments. Lambs spent more time consuming the milky flavor and the control at 123 and 144 s/test, respectively, compared with all other FCP (average of 65 s/test). In experiment 2, 12 weaned female Holstein Friesian calves (56-68 d of age) averaging 75.8 kg (standard deviation = 8.45) of body weight were offered a choice of 4 FCP (vanilla, milky, spicy/fenugreek, and red summer fruits) at an inclusion rate of 150 to 200 g/t (as-is basis) and the unflavored control for 5 min 4 times over 10 d. The average consumption rate was 27.8 g/min, and there were no differences among FCP. In experiment 3, a choice of 4 FCP with 2 different flavor combinations (vanilla-fenugreek and milky-vanilla) included at 75 g/t (as-is basis; low) or 150 g/t (high) was offered to a total of 12 weaned female Holstein Friesian calves (47-62 d of age) with an average body weight of 65.3 kg (standard deviation = 7.91). The FCP were offered daily for 14 d for 30 to 60 min/d. Vanilla-fenugreek (low) was consumed less at 57.5 g/test per calf compared with the other FCP (average of 87.5 g/test per calf). There were no other differences among FCP in experiment 3. Overall, compared with the control, flavors used in the present experiments did not affect feed intake of weaned lambs and calves. Other factors, such as taste, sight, texture of the feed, effect of the dams as previous experience (via maternal ingestion, which influences neonatal feeding), and their interactions, may also play a role in flavor preferences of young ruminants.

Evaluating internal and external markers versus fecal sampling procedure interactions when estimating intake in dairy cows consuming a corn silage-based diet.

Feed intake assessment is a valuable tool for herd management decisions. The use of markers, either internal or external, is currently the most used technique for estimating feed intake in production animals. The experiment used 10 multiparous Holstein cows fed a corn silage-based diet, with 55:45 forage-to-concentrate ratio, the average fecal recovery (FR) of TiO2 was higher than FR of Cr2O3, and both FR were more than unity. With internal markers, acetyl bromide lignin and cutin FR were lower than unity, and average FR for indigestible neutral detergent fiber (iNDF) and indigestible acid detergent fiber (iADF) was 1.5. The FR was unaffected by the fecal sampling procedure and appears to be an intrinsic property of each molecule and how it interacts with digesta. Of the 2 external markers, only Cr2O3 produced accurate fecal output (FO) estimates and the same happened to dry matter digestibility (DMD) when iNDF and iADF were used. Estimates for DMD and FO were affected by sampling procedure; 72-h bulk [sub-sample from total feces collection (TFC)] sampling consistently produced accurate results. The grab (sub-samples taken at specific times during the day) sampling procedures were accurate when using either of the indigestible fibers (iNDF or iADF) to estimate DMD. However, grab sampling procedures can only be recommended when concomitant TFC is performed on at least one animal per treatment to determine FR. Under these conditions, Cr2O3 is a suitable marker for estimating FO, and iNDF and iADF are adequate for estimating DMD. Moreover, the Cr2O3+iADF marker pair produces accurate dry matter intake estimates and deserves further attention in ruminant nutrition studies. The method of dosing the external markers is extremely important and greatly affects and determines results. Whichever the method, it must allow the animals to display normal feeding behavior and not affect performance. The grab sampling procedures can replace TFC (once FR is established), which may open new possibilities for pasture-based or collectively housed animals.

Systemic bone marker expression induced by grey and white mineral trioxide aggregate in normal and diabetic conditions.

AIM: To investigate the relationship between diabetes mellitus and local/systemic effects of both grey and white mineral trioxide aggregate (MTA) Angelus on bone marker expression. METHODOLOGY: Wistar rats were divided into two groups: healthy and diabetic (Alloxan induced), which were further divided into three subgroups (control, GMTA Angelus and WMTA Angelus). Polyethylene tubes filled with MTA materials or empty tubes were implanted in dorsal connective tissue. On days 7 and 30, blood samples were collected for calcium, phosphorus and ALP measurement. The animals were euthanized; implanted tubes were removed and processed for immunohistochemical analysis of osteocalcin (OCN) and osteopontin (OPN). Kruskal-Wallis followed by Dunn's multiple comparison test was performed for nonparametric data, and anova followed by Tukey's test for parametric data. RESULTS: No difference in systemic serum calcium levels between both groups was observed. On day 7, serum phosphorus levels within the WMTA healthy group were higher than that of the diabetic group. On day 30, healthy rats exhibited lower phosphorus levels than diabetic ones. At both time points, the diabetic group was associated with more ALP activity than the healthy group. Immunohistochemical analyses of the healthy group revealed OCN- and OPN-positive cells in the presence of both MTA materials. However, under diabetic conditions, both OCN and OPN were absent. CONCLUSION: Both MTA materials were associated with an increase in serum calcium, phosphorus and ALP, suggesting a potential systemic effect, along with triggered differentiation of OCN- and OPN-positive cells. Moreover, in diabetic conditions, an inhibitory effect on MTA-induced differentiation of OCN- and OPN-positive cells was detected.

RUNX-2, OPN and OCN expression induced by grey and white mineral trioxide aggregate in normal and hypertensive rats.

AIM: To investigate whether hypertension affects mineralization associated with white and grey mineral trioxide aggregate (MTA Angelus® ) implanted subcutaneously into rats by assaying osteoblastic biomarkers. METHODOLOGY: Polyethylene tubes containing grey MTA Angelus® , white MTA Angelus® , intermediate restorative material (IRM; positive control) or an empty tube (negative control) were implanted into the dorsal connective tissue of spontaneous hypertensive (n = 12) and Wistar (normotensive; n = 10) rats. Half of the rats in each group were killed after 7 days, and the remaining after 30 days. Tubes with surrounding tissue were removed, and immunostaining was performed to detect RUNX-2, OPN and OCN proteins. The normality of data was analysed using the Shapiro-Wilk test. Comparison of two independent groups was performed using the Mann-Whitney U-test, to detect a significant difference. A post hoc test accounting for multiple comparisons was performed following Tukey's test (P < 0.05). RESULTS: Under hypertensive conditions after 30 days, both MTA materials were associated with immunolabelling for RUNX-2 from low to moderate, which was less than that observed at normal blood pressure and the 7-day groups (P < 0.05). The expression of OPN and OCN proteins under both MTA conditions was considered low after both 7 and 30 days for the hypertensive condition, and was less than that in animals with normal blood pressure after 30 days (P < 0.05). No immunostaining for any biomarkers in the control and IRM groups was observed (P < 0.05). CONCLUSION: Hypertension decreased the immunostaining of RUNX-2, OPN and OCN biomarkers in response to MTA. Thus, hypertension can jeopardize the mineralization ability of MTA and may have a negative impact on endodontic treatment outcomes.

Efficacy of a high free iodine barrier teat disinfectant for the prevention of naturally occurring new intramammary infections and clinical mastitis in dairy cows.

Using a natural exposure trial design, the goal of our study was to evaluate the clinical efficacy of an iodine teat disinfectant with barrier properties and a high level of free iodine relative to a conventional iodine teat disinfectant with no barrier properties and low levels of free iodine. During the 18 wk of the trial, quarter milk samples were collected every 2 wk from 385 dairy cows from 2 herds. Cows on both farms were assigned in a balanced way according to milk yield, number of lactation, days in milk, somatic cell count (SCC) and microbiology culture pretrial into one of following groups: nonbarrier post milking teat disinfectant (NBAR; n = 195 cows; 747 quarters) or barrier postmilking teat disinfectant (BAR; n = 190 cows; 728 quarters). Afterward, at each scoring date every 2 wk, milk SCC was quantified in samples from all mammary quarters and microbiologic culture was only performed on milk samples with SCC >200,000 cells/mL for multiparous cows and SCC >100,000 cells/mL for primiparous cows. A new intramammary infection (NIMI) was defined when a quarter had milk SCC <200,000 cells/mL for multiparous cows and <100,000 cells/mL for primiparous without microorganism isolation, and in a subsequent sampling visit had milk SCC >200,000 cells/mL for multiparous cows and >100,000 cells/mL for primiparous cows, and positive microorganism isolation. A quarter could have several NIMI, but only 1 case per specific pathogen was considered. The most frequently isolated microorganism group on both farms was Streptococcus spp. (6.25% of total mammary quarters), followed by coagulase-negative staphylococci (3.6%) and Corynebacterium spp. (1.5%). In the present study, an interaction occurred between treatment and week of trial on the incidence risk of NIMI. Quarters disinfected with BAR had 54 and 37% lower odds of NIMI than quarters disinfected with NBAR at 8 and 16 wk of the trial, respectively; whereas at other weeks of the study both products had similar incidence risks of NIMI. Overall, teats disinfected with BAR had 46% lower odds of acquiring a clinical mastitis than those disinfected with NBAR. We concluded that the postmilking teat disinfectant with barrier properties and higher free iodine content reduced the risk of clinical mastitis, although differences in new infections were detected at only weekly time points.

Frequency of Chlamydia trachomatis infection in cervical intraepithelial lesions and the status of cytological p16/Ki-67 dual-staining.

BACKGROUND: Chlamydia trachomatis (Ct) is not a disease subject to mandatory reporting in Brazil, and the prevalence rate of this genital infection varies according to the region in which studies are conducted, as well as by the detection technique employed. Ct has been associated with persistence of Human papillomavirus (HPV) infection and the facilitation of cervical carcinoma development. We evaluated the Chlamydia trachomatis infection and its association with cytology, p16/Ki-67 dual-stained cytology and cervical intraepithelial lesions status in a screening cohort in Brazil. METHODS: This was a cross-sectional study of 1481 cervical samples from asymptomatic women aged 18 to 64. Samples were collected for liquid-based cytology and Ct detection by polymerase chain reaction. p16/Ki-67 double staining was performed on samples with abnormal cytology. Statistical analysis was by chi-square and likelihood-ratio tests. Odds ratio (OR) and 95% confidence intervals (95% CI) were determined. RESULTS: The frequency of Ct was 15.6% and its presence was not associated with detection of p16/Ki-67 [OR = 1.35 (0.5-3.4)]. There was also no association between abnormal cervical cytology and Ct-positivity [OR = 1.21 (0.46-3.2)]. Associations were observed between p16/Ki-67 and high-grade lesions detected by cytology and in biopsies [OR = 3.55 (1.50-8.42) and OR = 19.00 (0.6-7.2), respectively]. CONCLUSIONS: The asymptomatic women in our study had a high frequency of Ct infection but this was not associated with p16/Ki-67 detection in samples with abnormal cytology. The expression of p16/Ki-67 was highest in women with high-grade CIN (p = 0.003).

Copper accumulation by stickleback nests containing spiggin.

The three-spined stickleback is a ubiquitous fish of marine, brackish and freshwater ecosystems across the Northern hemisphere that presents intermediate sensitivity to copper. Male sticklebacks display a range of elaborate reproductive behaviours that include nest construction. To build the nests, each male binds nesting material together using an endogenous glycoprotein nesting glue, known as 'spiggin'. Spiggin is a cysteine-rich protein and, therefore, potentially binds heavy metals present in the environment. The aim of this study was to investigate the capacity of stickleback nests to accumulate copper from environmental sources. Newly built nests, constructed by male fish from polyester threads in laboratory aquaria, were immersed in copper solutions ranging in concentration from 21.1-626.6 µg Cu L(-1). Bundles of polyester threads from aquaria without male fish were also immersed in the same copper solutions. After immersion, nests presented higher amounts of copper than the thread bundles, indicating a higher capacity of nests to bind this metal. A significant, positive correlation between the concentration of copper in the exposure solution and in the exposed nests was identified, but there was no such relationship for thread bundles. Since both spiggin synthesis and male courtship behaviour are under the control of circulating androgens, we predicted that males with high courtship scores would produce and secrete high levels of the spiggin protein. In the present study, nests built by high courtship score males accumulated more copper than those built by low courtship score males. Considering the potential of spiggin to bind metals, the positive relationship between fish courtship and spiggin secretion seems to explain the higher amount of copper on the nests from the fish showing high behaviour scores. Further work is now needed to determine the consequences of the copper binding potential of spiggin in stickleback nests for the health and survival of developing embryos.

Effect of dietary cation-anion difference on ruminal metabolism, total apparent digestibility, blood and renal acid-base regulation in lactating dairy cows.

The present study aimed to evaluate the effect of dietary cation-anion difference (DCAD) on ruminal fermentation, total apparent digestibility, blood and renal metabolism of lactating dairy cows. Sixteen Holstein cows were distributed in four contemporary 4×4 Latin Square designs, which consisted of four periods of 21 days and four treatments according to DCAD: +290; +192; +98 and -71 milliequivalent (mEq)/kg dry matter (DM). Ruminal pH and concentrations of acetic and butyric acid increased linearly according to the increase of DCAD. Similarly, NDF total apparent digestibility linearly increased by 6.38% when DCAD increased from -71 to 290 mEq/kg DM [Y=65.90 (SE=2.37)+0.0167 (SE=0.0068)×DCAD (mEq/kg DM)]. Blood pH was also increased according to DCAD, which resulted in reduction of serum concentrations of Na, K and ionic calcium (iCa). To maintain the blood acid-base homeostasis, renal metabolism played an important role in controlling serum concentrations of Na and K, since the Na and K urinary excretion increased linearly by 89.69% and 46.06%, respectively, from -71 to 290 mEq/kg DM. Changes in acid-base balance of biological fluids may directly affect the mineral composition of milk, as milk concentrations of Na, K, iCa and chlorides were reduced according to blood pH increased. Thus, it can be concluded that the increase of DCAD raises the pH of ruminal fluid, NDF total apparent digestibility, and blood pH, and decreases the milk concentration of cationic minerals, as well as the efficiency of Na utilization to milk production.

Effect of dietary cation-anion difference on performance of lactating dairy cows and stability of milk proteins.

Casein micelle stability is negatively correlated with milk concentrations of ionic calcium, which may change according to the metabolic and nutritional status of dairy cows. The present study aimed to evaluate the effect of dietary cation-anion difference (DCAD) on concentrations of casein subunits, whey proteins, ionic calcium, and milk heat and ethanol stability. Sixteen Holstein cows were distributed in 4 contemporary 4 × 4 Latin square designs, which consisted of 4 periods of 21 d and 4 treatments according to DCAD: 290, 192, 98, and -71 mEq/kg of dry matter (DM). The milk concentrations of ionic calcium and κ-casein were reduced as DCAD increased, whereas the milk urea nitrogen and ß-lactoglobulin concentrations were increased. As a result of these alterations, the milk ethanol stability and milk stability during heating at 140 °C were increased linearly with increasing DCAD [Y = 74.87 (standard error = 0.87) + 0.01174 (standard error = 0.0025) × DCAD (mEq/kg of DM) and Y = 3.95 (standard error = 1.02) + 0.01234 (standard error = 0.0032) × DCAD (mEq/kg of DM), respectively]. In addition, 3.5% fat-corrected milk and fat, lactose, and total milk solids contents were linearly increased by 13.52, 8.78, 2.5, and 2.6%, respectively, according to DCAD increases from -71 to 290 mEq/kg of DM, whereas crude protein and casein content were linearly reduced by 4.83 and 4.49%, respectively. In conclusion, control of metabolic changes in lactating dairy cows to maintain blood acid-base equilibrium plays an important role in keeping milk stable to ethanol and during heat treatments.

The effect of timing of the induction of ovulation on embryo production in superstimulated lactating Holstein cows undergoing fixed-time artificial insemination.

Two experiments evaluated the effects of timing of the induction of ovulation in superstimulated lactating Holstein donor cows that were fixed-time artificially inseminated. Secondary objectives were to evaluate the effects of the timing of progesterone (P4) device removal (Experiment 1) or the addition of a second norgestomet implant (Experiment 2) during superstimulation. In Experiment 1, 12 cows were allocated to one of four treatment groups with the timing of P4 device removal (24 or 36 h) and pLH treatment (48 or 60 h), after the first PGF as main factors, in a Latin Square (cross-over) design. There was an interaction (P = 0.03) between time of P4 device removal and time of pLH treatment. Mean (± SEM) numbers of transferable embryos were higher when the P4 device was removed at 36 h and pLH was administered at 60 h after the first PGF (P36LH60 =6.3 ± 1.4) compared to other treatments (P24LH60 =3.7 ± 1.1; P24LH48 =2.4 ± 0.8; or P36LH48 =2.2 ± 0.7). In Experiment 2, 40 cows were randomly allocated into one of four treatments with the number of norgestomet implants (one or two) and the time of induction of ovulation with GnRH relative to the first PGF (48 vs. 60 h) as main effects. The mean number of transferable embryos was higher (P = 0.02) when GnRH was administered at 60 h (4.2 ± 1.3) compared to at 48 h (2.7 ± 0.8), and the number of freezable embryos was increased (P = 0.01) in cows receiving two (3.0 ± 1.0) rather than one norgestomet implant (1.5 ± 0.5). In summary, embryo production in lactating Holstein cows was increased when the ovulatory stimulus (pLH or GnRH) was given 60 h after the first PGF, particularly when the P4 device was removed 36 h after the first PGF and when two norgestomet ear implants were used during the superstimulation protocol.

Timing of insemination using sex-sorted sperm in embryo production with Bos indicus and Bos taurus superovulated donors.

Two experiments were designed to evaluate the effect of different insemination times (12 and 24h or 18 and 30h) and different types of semen (sex-sorted or non-sorted sperm) on embryo production in Nelore (Bos indicus) and Holstein (Bos taurus) superstimulated donors. In the first experiment, hormonal superstimulation of ovarian follicular development in Nelore donors (n=71) was performed in randomly allocated animals to one of the three treatment groups, and they were inseminated at 12 and 24h after an ovulatory stimulus with pLH treatment was applied, either with sex-sorted (4.2×10(6) sperm/insemination; S12/24; n=17) or non-sorted sperm (20×10(6) sperm/insemination; NS12/24; n=18), or they were inseminated at 18 and 30h using sex-sorted sperm (4.2×10(6) sperm/insemination; S18/30; n=19). A greater number of transferable embryos were found when sex-sorted sperm was used to inseminate the animals at 18 and 30h (4.5±3.0) compared to insemination at 12 and 24h (2.4±1.8; P<0.001). However, a greater embryo production (6.8±2.6) was obtained with non-sorted sperm. In the second experiment, the same insemination times and semen types were used in lactating high-production Holstein cows (n=12). A crossover design was employed in this trial. A lesser embryo production (P=0.007) was found in Holstein donors that were inseminated using sex-sorted sperm at 12 and 24h (4.6±3.0) compared to non-sorted sperm (8.7±2.8). However, intermediate results were obtained when the inseminations with sex-sorted sperm were performed at 18 and 30h (6.4±3.1). These results supported the current hypothesis that it is possible to improve embryo production using sex-sorted sperm in B. indicus and B. taurus superstimulated donors when the inseminations are performed near the same time as time-synchronized ovulations. However, the embryo production for timed artificial insemination (TAI) with sex-sorted sperm was still less than the production with non-sorted sperm.

Timed embryo transfer programs for management of donor and recipient cattle.

Currently, timed ovulation induction and fixed-time artificial insemination (FTAI) in superstimulated donors and synchronization protocols for fixed-time embryo transfer (FTET) in recipients can be performed using GnRH or estradiol plus progesterone/progestin (P4)-releasing devices and prostaglandin F(2α) (PGF2α). The control of follicular wave emergence and ovulation at predetermined times, without estrus detection, has facilitated donor and recipient management. However, because Bos taurus cows have subtle differences in their reproductive physiology compared with Bos indicus cattle, one cannot assume that similar responses will be achieved. The present review will focus on the importance of orchestrating donor and recipient management to assure better logistics of procedures to achieve more desirable results with embryo collection and transfer. In addition, this will provide clear evidence that the use of FTAI in superstimulated donors and FTET in embryo recipients eliminates the need to detect estrus with satisfactory results. These self-appointed programs reduce labor and animal handling, facilitating the use of embryo transfer in beef and dairy cattle.

Sustained efficacy and immunogenicity of the HPV-16/18 AS04-adjuvanted vaccine up to 7.3 years in young adult women.

We report efficacy and immunogenicity of the HPV-16/18 AS04-adjuvanted vaccine up to 7.3 years post-vaccination. The study was conducted in a population (N=433) of women enrolled in Brazilian centres from an initial placebo-controlled study. Women were aged 15-25 years at first vaccination. During the most recent year of follow-up, approximately 7 years after initial vaccination, no cases of infection or cytohistological lesions associated with HPV-16/18 were observed in the vaccinees. Vaccine efficacy (95% confidence interval) up to 7.3 years was 94.5% (82.9, 98.9) for incident infection, 100% (55.7, 100) for 12-month persistent infection and 100% (-129.8, 100) for cervical intraepithelial neoplasia grade 2+. Antibody titres for total IgG and neutralising antibodies remained several folds above natural infection levels and >or=96% of women were seropositive. Vaccine safety was similar to placebo. This is the longest follow-up study for a licensed cervical cancer vaccine.

Sustained efficacy and immunogenicity of the human papillomavirus (HPV)-16/18 AS04-adjuvanted vaccine: analysis of a randomised placebo-controlled trial up to 6.4 years.

BACKGROUND: Prophylactic human papillomavirus (HPV) vaccines have to provide sustained protection. We assessed efficacy, immunogenicity, and safety of the HPV-16/18 AS04-adjuvanted vaccine up to 6.4 years. METHODS: Women aged 15-25 years, with normal cervical cytology, who were HPV-16/18 seronegative and oncogenic HPV DNA-negative (14 types) at screening participated in a double-blind, randomised, placebo-controlled initial study (n=1113; 560 vaccine group vs 553 placebo group) and follow-up study (n=776; 393 vs 383). 27 sites in three countries participated in the follow-up study. Cervical samples were tested every 6 months for HPV DNA. Management of abnormal cytologies was prespecified, and HPV-16/18 antibody titres were assessed. The primary objective was to assess long-term vaccine efficacy in the prevention of incident cervical infection with HPV 16 or HPV 18, or both. We report the analyses up to 6.4 years of this follow-up study and combined with the initial study. For the primary endpoint, the efficacy analysis was done in the according-to-protocol (ATP) cohort; the analysis of cervical intraepithelial neoplasia grade 2 and above (CIN2+) was done in the total vaccinated cohort (TVC). The study is registered with ClinicalTrials.gov, number NCT00120848. FINDINGS: For the combined analysis of the initial and follow-up studies, the ATP efficacy cohort included 465 women in the vaccine group and 454 in the placebo group; the TVC included 560 women in the vaccine group and 553 in the placebo group. Vaccine efficacy against incident infection with HPV 16/18 was 95.3% (95% CI 87.4-98.7) and against 12-month persistent infection was 100% (81.8-100). Vaccine efficacy against CIN2+ was 100% (51.3-100) for lesions associated with HPV-16/18 and 71.9% (20.6-91.9) for lesions independent of HPV DNA. Antibody concentrations by ELISA remained 12-fold or more higher than after natural infection (both antigens). Safety outcomes were similar between groups: during the follow-up study, 30 (8%) participants reported a serious adverse event in the vaccine group versus 37 (10%) in the placebo group. None was judged related or possibly related to vaccination, and no deaths occurred. INTERPRETATION: Our findings show excellent long-term efficacy, high and sustained immunogenicity, and favourable safety of the HPV-16/18 AS04-adjuvanted vaccine up to 6.4 years. FUNDING: GlaxoSmithKline Biologicals (Belgium).