2 mM exogenous FFA decreased the abundance of MFN2 and upregulated phosphorylation levels of the inhibitor of NF-κB (IκB) α and NF-κB p65, the IκB kinase β activity, and the abundance of NLRP3, caspase-1, IL-1β, IL-6, and TNF-α. Whereas MFN2 knockdown potentiated the FFA-induced activation of these inflammatory pathways, overexpression of MFN2 attenuated the detrimental effect of excess exogenous FFA by improving mitochondrial function and decreasing the release of reactive oxygen species, suggesting that MFN2 may be a potential therapeutic target for FFA-induced hepatic inflammation in dairy cows during early lactation. The objective of this study was to determine the effects of adding molasses-based liquid feed to a high-straw dry cow diet on intake, behavior, health, and performance of dairy cows across the transition period. Holstein cows (n = 40) entering their second lactation or greater were enrolled at dry-off (~45 d before expected calving) and assigned to 1 of 2 treatments a high-straw dry cow total mixed ration (TMR) with either (1) no molasses [CON; n = 20; 10.7% crude protein (CP), 1.45 Mcal/kg of net energy for lactation (NEL)] or (2) supplemented with 1 kg/cow daily [dry matter (DM) basis] of molasses-based liquid feed (LF; n = 20; 11.2% CP, 1.49 Mcal/kg of NEL). At calving all cows were fed the same lactating TMR (14.9% CP, 1.68 Mcal/kg of NEL) and followed for 28 d. During the dry period, cows fed the LF diet had greater DM intake (15.3 ± 0.24 vs. 13.4 ± 0.24 kg/d) and tended to have a shorter interval between meals (192.8 ± 8.38 vs. 216.5 ± 7.97 min) and more frequent meals (6.3 ± 0.27 vs. 5.6 ± 0.27 meals/draw dry cow diets may improve intake and consistency in nutrients consumed during the dry period and in early lactation, as well as possibly promoting better rumen health across the transition period. 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. The objective of this study was to evaluate different analytical methods of assessing failure of passive transfer (FPT) in neonatal calves. We hypothesized that 3 different media (i.e., centrifuged serum, centrifuged plasma, filtered plasma) and different analytical methods [i.e., ELISA, capillary electrophoresis (CE), Brix refractometer, and handheld optical refractometer] would be highly correlated with the gold standard radial immunodiffusion (RID) and would generate comparable results. Serum and plasma blood samples were collected from Holstein Friesian calves (n = 216) aged 1 to 7 d, from 2 commercial dairy herds in northeast Germany. The RID analysis showed that 59 of 216 calves (27%) had serum IgG concentrations of less then 10 mg/mL and 157 calves (73%) had serum concentrations of ≥10 mg/mL. The mean IgG concentration (± standard deviation) was 17.1 ± 9.8 mg/mL, and the range was 0.8 to 47.8 mg/mL. In serum, the correlation between RID and CE was r = 0.97, and between RID and ELISA was r = 0.90; CE and ELISA were also highly correlated (r = 0.89). Both refractometry methods were highly correlated with RID using centrifuged serum, centrifuged plasma, or filtered plasma (Brix refractometer r = 0.84, 0.80, and 0.78, respectively; handheld optical refractometer r = 0.83, 0.81, and 0.80, respectively). We determined test characteristics (optimum thresholds, sensitivity, specificity, positive predictive value, negative predictive value, and area under the curve) for CE, ELISA, and the handheld optical and digital refractometers using receiver operating characteristic curve analyses with RID as the reference value. Optimal thresholds for assessing FPT using plasma were higher than for serum, regardless of the method of plasma harvesting. The 4 different devices had comparable areas under the curve, irrespective of the medium used. All analytical methods can be used to assess FPT. The objectives of this study were to (1) describe the calving location of dairy cattle given access to a pasture and barn; (2) identify factors associated with calving location; and (3) compare the lying and exploratory behavior of cows in the 24 h before calving and a previous day. Seventy-two Holstein dairy heifers and cows (n = 36 nulliparous and n = 36 primiparous and multiparous combined) were housed in a covered bedded-pack barn (167.4 m2) with free access to 2.1 ha of pasture. The composition of the group was dynamic, because cows were moved in weekly at 19 ± 6 d [mean ± standard deviation (SD)] before their calving date, and were removed immediately after calving. To facilitate data collection, we divided the environment into 9 sections, including the barn (section 1; 167.4 m2), 7 sections of open pasture (sections 2 to 8; 2,402 ± 60 m2), and 1 section of pasture surrounded by natural forage cover (section 9; 3,593 m2). We then collapsed these 9 sections into 3 distinct areas for further analysis the heat stress was low (THI ≤68) selected the area with natural forage cover more frequently than the barn. On the calving day, cows spent more time lying down with more short bouts of lying, and crossed more sections compared with the baseline day. Steps were affected by an interactive effect of day, parity, and heat stress; nulliparous heifers took more steps on the calving day during conditions of heat stress compared with no heat stress. Results indicate that cows and heifers had different preferences for their environment at calving, and when provided access to pasture, both changed their lying and exploratory behavior on the day of calving compared with a previous day. Functional traits, such as fertility and lactation persistency, are becoming relevant breeding goals for dairy cattle. Fertility is a key element for herd profitability and animal welfare; in particular, calving interval (CIN) is an indicator of female fertility that can be easily recorded. Lactation persistency (LPE; i.e., the ability of a cow to maintain a high milk yield after the lactation peak) is economically important and is related to several other traits, such as feed efficiency, health, and reproduction. The selection of these functional traits is constrained by their low heritability. In this study, variance components for CIN and LPE in the Italian Simmental cattle breed were estimated using genomic and pedigree information under the single-step genomic framework. A data set of 594,257 CIN records (from 275,399 cows) and 285,213 LPE records (from 1563,389 cows) was considered. Phenotypes were limited up to the third parity. The pedigree contained about 2 million animals, and 7,246 genotypes were ae Italian Simmental breeding program. The use of a single-step approach may provide better results for young genotyped animals without their own phenotypes. The present review focuses on methyl donor metabolism and nutrition in the periparturient and lactating dairy cow. Methyl donors are involved in one-carbon metabolism, which includes the folate and Met cycles. These cycles work in unison to support lipid, nucleotide, and protein synthesis, as well as methylation reactions and the maintenance of redox status. A key feature of one-carbon metabolism is the multi-step conversion of tetrahydrofolate to 5-methyltetrahyrofolate. Homocysteine and 5-methyltetrahyrofolate are utilized by vitamin B12-dependent Met synthase to couple the folate and Met cycles and generate Met. Methionine may also be remethylated from choline-derived betaine under the action of betaine hydroxymethyltransferase. Regardless, Met is converted within the Met cycle to S-adenosylmethionine, which is universally utilized in methyl-group transfer reactions including the synthesis of phosphatidylcholine. Homocysteine may also enter the transsulfuration pathway to generate glutathione or taurine fontation, fatty acid feeding strategies that may optimize methyl donor supplementation efficacy, and potential epigenetic mechanisms are also considered. Milk production and time effects are considered related to heat stress but they have not yet been combined in predictive models. In two parts, this study aimed to develop new models to predict heat stress (rectal temperature and respiration rate) of lactating dairy cows by inputting predictors, including ambient temperature (Ta), relative humidity (RH), wind speed (WS), milk yield (MY), and time blocks. In the first part of the study, we built the quantitative foundation for the second part, including the regression relation between respiration rate and rectal temperature (to convert predicted respiration rate to predicted body temperature), as well as between rectal temperature and respiration rate when heat stress was triggered (to recognize whether herds were under stress). In the second part, we built models that combined the abovementioned predictors to predict respiration rate. In part I, data were obtained from 45 high-producing Holstein cows within a Ta range of 9.5 to 30.8°C. We found a very strong c30-0800 h, respectively (reducing the intercept of the expression in 0630-0800 h). Compared with temperature-humidity index equations, the proposed model performed better at suppressing prediction error, and had better sensitivity and accuracy in recognizing whether heat stress was triggered. Given consumer interest in Mozzarella di latte di Bufala and other cheeses, and the growing interest of the cheese industry in offering products adequate for lactovegetarian consumers, this study aimed to compare clotting capacity of vegetal and animal rennet in buffalo milk. Milk coagulation properties of 1,261 buffalo bulk milk samples collected during milk quality testing were assessed by lactodynamography using commercial animal (75% chymosin and 25% bovine pepsin) and vegetal (Cynara cardunculus) rennets. Chemical composition of milk samples was predicted by MilkoScan (Foss Analytics, Hillerød, Denmark) calibrated with specific buffalo standards. Rennet effect (animal versus vegetal) was statistically analyzed with a paired t-test. Fat, protein, and lactose contents of milk samples were 7.94%, 4.52%, and 4.80%, respectively. A similar variability of milk coagulation properties was observed with both rennets, with the exception of greater variability of curd firmness at 30 min after the addition of vegetal rennet compared with animal rennet (73 and 26%, respectively).