Mid-lactation Cows Research Articles

Potential involvement of PPARs in the inhibition of mammary lipid synthesis during diet-induced milk fat depression

The objective of this study was to evaluate the possible role of the peroxisome proliferator-activated receptors (PPARs: PPAR-α, PPAR-β/δ, and PPAR-γ) in diet and conjugated linoleic acid (CLA)-induced milk fat depression (MFD) in dairy cows. We hypothesized that the expression of PPARs, which regulate lipid metabolism and bind to polyunsaturated fatty acids, could be modulated by biohydrogenation intermediates that induce MFD, thereby interfering with milk fat synthesis. First, tissue profiling revealed that PPAR-α and PPAR-β/δ had low expression in mammary tissue compared with the liver. A comparison of lactating and nonlactating tissue from the same cows showed that expression of the PPARs did increase during lactation. Mammary expression of the PPARs during MFD was then observed in 9 mid-lactation cows in a 3x3 Latin square design with MFD induced by a 3-d intravenous infusion of trans-10,cis-12 CLA or feeding a high-oil/low-forage diet. The expression of the PPARs remained largely unaltered during CLA and diet-induced MFD, except for an increase in PPAR-α target genes CPT1A and ACADVL that are involved in β-oxidation. The interaction of PPAR-γ chemical agonist Troglitazone and antagonist T0070907 and CLA was then investigated in bovine mammary epithelial cells (MAC-T). The activation and inhibition of PPAR-γ did not overcome trans-10,cis-12 CLA inhibition of lipogenesis despite the agonist stimulating PPAR-γ expression. Furthermore, PPAR-γ activation did not modify the expression of lipogenic genes. Overall, the results fail to support a functional role of the PPARs in the inhibition of lipogenesis during MFD in dairy cows.

Validation of a Remote Sampling Sensor for Measuring Urine Volume and Nitrogen Concentration in Grazing Dairy Cattle.

The purpose of this research was to validate a urine sensor (Lincoln University PEETER V2.0, Canterbury, New Zealand) that records the time and volume of urination events for dairy cows in addition to collecting a proportional urine sample from all urination events. Sixteen multiparous Holstein × Jersey mid-lactating cows (101 ± 5 days in milk, 498 ± 24.2 kg body weight, 26.2 ± 3.07 kg/d milk yield; mean ± standard deviation) were allocated herbage diets ranging in protein and sodium content to generate a range of urine volumes and urine nitrogen (UN) concentrations. Total collection of individual urination events occurred during a 72-h measurement period where PEETER V2.0 sensors were attached to cows. A mixed model ANOVA using lme4 package (version 1.1-35.5) in R (version 4.3.3) were used to compare the means. The average urine event size was 2.65 ± 1.1 L for total collection by observers and 2.68 ± 1.1 L as recorded by the sensor (mean ± standard deviation; p = 0.730). The urine nitrogen concentration was 5.76 ± 1.2 g N/L for samples collected by observers and 5.85 ± 1.3 g N/L for the samples collected by the sensor (p = 0.583). The calculated UN excretion was 156 ± 45.1 g/day for direct measurements and 162 ± 40.0 g/day for the sensor (p = 0.539. Contrasts with simultaneously measured data were undertaken using Lin's Concordance Correlation Coefficient (CCC) and a Pearson correlation coefficient (r). Correlations between the actual values and sensor values were strong, with little to moderate variability in the urine volume (CCC = 0.936, r = 0.937; n = 222), UN concentration (CCC = 0.840, r = 0.837, n = 48) and total UN excretion (CCC = 0.827, r = 0.836, n = 24). Considering the findings, the PEETER V2.0 urine sensor has the potential to reliably measure urine volumes and UN concentrations for estimations of the UN excretion of dairy cattle under grazing systems.

517 The effects of pellet starch concentration and allocation amount for Holstein cows in peak, mid, and late lactation in an automatic milking system

Abstract This study evaluated the effects of pellet starch concentration and amount of pellet delivered in an automated milking system (AMS) on AMS pellet and partial mixed ration (PMR) intake, milk and milk component yield, and milking and feeding behavior for Holstein cows at different stages of lactation. Twenty-four Holstein cows at peak [n = 8; 85 ± 25.7 days in milk (DIM)], mid (n = 8; 185 ± 35.1 DIM) or late (n = 8; 290 ± 69.5 DIM) lactation (3 squares/DIM category), housed in a free-stall barn with a feed-first guided traffic flow AMS, were used. Treatments included low (LS; 24% DM) or high (HS; 34% DM) starch pellets that were provided at low (LA; 2 kg/d DM) or high (HA; 6 kg/d DM) quantities within replicated 4 × 4 Latin square design. Each period lasted 21 d including 16 d of adaptation and 5 d of data and sample collection to evaluate feed intake and behavior, milking characteristics, milk and milk component yields. Pellet starch did not affect the amount of pellet consumed, but HA cows consumed more pellet than LA (4.3 vs. 1.8 kg/d; P < 0.01). Relative to LA, HA had greater pellet refused in the AMS (P < 0.01) and decreased PMR intake (P = 0.04). Total DMI was 1.3 kg/d greater for HA than LA cows (P = 0.05). Pellet starch and DIM did not affect PMR intake or DMI. Neither pellet starch nor allocation affected the number of PMR meals; however, LA increased PMR eating time by 20 min/d (P < 0.01) and PMR meal length by 2 min/meal (P = 0.03). Milking frequency was not affected by pellet starch or DIM, but HA tended to increase total milking frequency (2.7 vs. 2.6 no/d; P = 0.06) over LA, with greater voluntary milkings (2.5 vs 2.3 no/d; P < 0.01). Milk yield/visit and milking duration were not affected by pellet starch or allocation. Milk yield was not affected by pellet starch, or the amount allocated averaging 43.2, 41.3 and 43.2 kg/d for peak, mid, and late lactation cows (P ≥ 0.19). Compared with LS, HS decreased milk fat concentration (4.1 vs 3.9%; P < 0.01) and providing the HA reduced milk fat concentration (4.1 vs 3.96%; P < 0.01) when compared with LA; however, neither pellet starch nor allocation affected fat yield, averaging 1.67 kg/d (P ≥ 0.15). True protein yield was not affected. Cows fed the LA had greater milk urea nitrogen (14.8 mg/dL) relative to HA (14.3 mg/dL; P = 0.03). Increasing the amount of pellet allocated in the AMS reduced PMR intake, while increasing total DMI and resulted in greater amounts of pellet refusals by cows; however, greater amounts of pellet may increase attendance at the AMS without affecting milk or milk component yields. Starch concentration of the pellet had little effect on productivity.

The effect of flow-responsive vacuum and pulsation with early attachment of milking unit on teat tissue condition and milking performance in Holstein dairy cows

The objectives of this study were to assess the effect of flow-responsive vacuum and pulsation, in conjunction with the early attachment of the milking unit (TRT), on teat tissue conditions and milking characteristics in dairy cows. In a switch-back trial, 5,235 Holstein cows milked 3 times daily in a rotary parlor were assigned to the TRT or control (CON) group. The trial lasted 84 d and was comprised of 4 alternating 3-week periods of TRT and CON. For both groups, premilking udder preparation consisted of teat brushing, forestripping and predipping, and wiping of teats, resulting in a stimulation time of 4 s. In the TRT group, the preparation lag time was 58 s, and in the CON group, it was 91 s for early- and mid-lactation cows and 105 s for late-lactation animals. Upon milking unit attachment, the TRT cows were milked at a lower vacuum (37.6 kPa) and pulsation (50 cycles/min, pulsation ratio of 30:70). The vacuum and pulsation settings were changed to milking mode when the milk flow reached 0.5 kg/min (pulsation switch-point) and 1.6 kg/min (vacuum switch-point). For milking mode, the vacuum setting was 47.7 kPa, and the pulsation rate was 60 cycles/min at a ratio of 65:35. The CON cows were milked with a flow-responsive vacuum, using the same vacuum settings as the TRT group. We assessed machine milking-induced short-term teat tissue changes and teat-end hyperkeratosis by palpation and visual inspection postmilking. Electronic on-farm milk meters were used to assess milking characteristics. Generalized linear mixed models were used to analyze the effect of treatment on the outcome variables. Compared with cows in group CON, the odds ratios (95% confidence interval; 95% CI) of short-term teat-tissue changes in early-, mid-, and late lactation cows in group TRT were 0.62 (0.52-0.76), 0.61 (0.48-0.77), and 0.93 (0.76-1.14), respectively. The least squares means [LSM, (95% CI)] for milking unit-on time in early-, mid-, and late lactation animals, respectively, were 251 (248-253), 236 (234-238), and 220 (218-222) s for group TRT and 247 (245-249), 232 (230-234), and 214 (213-216) s for the CON group. The LSM (95% CI) of peak milk flow rate in early-, mid-, and late lactation animals, respectively, were 5.75 (5.68-5.82), 5.77 (5.70-5.84), and 5.54 (5.48-5.59) kg/min for the TRT cows and 5.65 (5.58-5.72), 5.74 (5.68-5.81), and 5.45 (5.40-5.51) kg/min for the CON cows. The odds ratios (95% CI) of forced take-off in group TRT for early-, mid-, and late lactation cows, respectively, were 0.39 (0.37-0.41), 0.32 (0.30-0.34), and 0.47 (0.44-0.52) compared with their respective CON groups. In this study, cows that were milked using flow-responsive vacuum and pulsation with early attachment of the milking unit had lower odds of short-term teat tissue changes and forced take-off, as well as a higher peak milk flow rate. Our data suggest that the application of flow-responsive vacuum and pulsation facilitates early attachment of the milking unit, improves teat tissue condition, and has the potential to improve parlor efficiency.

Effects of brewery by-products based silage on productive performance of crossbred dairy cows.

The present study aimed to evaluate the effect of an increasing levels of brewery by-products based silage on productive performances of 3/4 Friesian x Boran mid-lactating cows. Experimental cows had similar in initial milk yield (11.7 ± 1.0), average days in milk (81.7 ± 6.1) and live weight (LW, 430.7 ± 40.3kg) but different in parities (2-5).The dietary treatments were arranged randomly in 4 × 4 Latin Square Design that included ad libitum natural pasture hay feeding for all treatments as a roughage source plus a commercial dairy concentrate mix supplemented at 0.5kg DM (dry matter)/liter of milk produced/day for cows in the control group (T1) and 0.3, 0.5 and 0.7kg DM of brewery by-products based silage per liter of milk yield/cow/day for cows in T2, T3 and T4 groups, respectively. The study revealed that the daily milk yield of experimental cows was influenced by dietary treatments with relatively higher daily milk yield being recorded (P < 0.05) for cows in the T4 (13.9l) followed by T3 (13.8l). Milk composition of cows remained unchanged (P > 0.05) except for fat percentage of the milk that showed a declining trend (P < 0.05) with incremental inclusion levels of brewery by-products based silages. The highest net income (NI, 437.9 Eth. Birr) and marginal rate of return (MRR, 800.7%) was obtained for cows receiving brewery by-products based silage at the rate of 0.7kg/liter of milk yield as compared to cows in the other treatment groups. Further study is required on the long term effect of brewery by-products based silage supplementation on productive, reproductive performance, and milk microbial qualities.

Impact of milking interval and time on milk spontaneous lipolysis and composition in dairy cows

Milk lipolysis is defined as the hydrolysis of triglycerides, the major component of milk fat, resulting in the release of short-chain fatty acids (FA) responsible for rancid flavor and partial glycerides that impair functional properties such as foaming and creaming abilities. Milk lipolysis is a complex phenomenon that depends on both animal parameters and breeding factors. Milk spontaneous lipolysis is known to be higher in milk from evening milkings than from morning milkings. This may be related to the longer length of overnight milking intervals or to the nycthemeral cycle. In this experiment, our objective was thus to study the impact of both milking intervals and time of day on milk spontaneous lipolysis in twice-daily-milking systems with one of 3 milking intervals: Short Day - Long Night (SD-LN, 6.30 a.m. and 4.30 p.m.,); Long Day - Short Night (LD-SN, 6:30 a.m. and 8:30 p.m.,); and Balanced Day and Night (BDN, 6:30 a.m. and 6:30 p.m.,). To achieve this goal, 21 multiparous dairy cows in mid-lactation were used in a 3 × 3 Latin square design over 3 periods. The experiment lasted 5 weeks, corresponding to 3 experimental periods of 6 d alternating with 8 d of milking with conventional hours (morning-evening gap of 10 h). We confirmed that milk spontaneous lipolysis was influenced by milking interval, but not the milking time. Indeed, we observed more lipolysis in SD-LN evening milk (+0.20 mEq/100 g fat) and LD-SN morning milk (+0.22 mEq/100 g fat), both of which corresponded to a 10 h interval between successive milkings. High lipolysis milk came from cows that produced less milk with a higher milk fat content. No significant difference between milkings was observed for BDN. Milk protein, total P and citrate contents increased according to the duration of mammary gland storage of milk (from 10 to 14 h). There was no effect of milking intervals on milk fat globule diameter. The milk Na+/K+ ratio, indicating an opening of tight junctions in the mammary gland, increased only in evening milkings with BDN and LD-SN. In conclusion, we found that the effect of milking intervals on lipolysis is stronger than that of the nycthemeral cycle.

Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS

This study evaluated the effects of supplementing the diet of lactating cows with Acremonium terrestris culture (ATC) on milk production, serum antioxidant capacity, inflammatory indices, and serum lipid metabolomics. Over 90 days, 24 multiparous Chinese Holstein cows in mid-lactation (108 ± 10.4 days in milk, 637 ± 25 kg body weight, 30.23 ± 3.7 kg/d milk yield) were divided into either a control diet (CON) or a diet supplemented with 30 g of ATC daily. All the data were analyzed using Student’s t test with SPSS 20.0 software. The results showed that compared with CON feeding, ATC feeding significantly increased milk yield, antioxidant capacity, and immune function. Lipidome screening identified 143 lipid metabolites that differed between the two groups. Further analysis using “random forest” machine learning revealed three glycerophospholipid serum metabolites that could serve as lipid markers with a predictive accuracy of 91.67%. This study suggests that ATC can be a useful dietary supplement for improving lactational performance in dairy cows and provides valuable insights into developing nutritional strategies to maintain metabolic homeostasis in ruminants.

Epigallocatechin-3-gallate protects bovine ruminal epithelial cells against lipopolysaccharide-induced inflammatory damage by activating autophagy

BackgroundSubacute ruminal acidosis (SARA) causes an increase in endotoxin, which can induce immune and inflammatory responses in the ruminal epithelium of dairy cows. In non-ruminants, epigallocatechin-3-gallate (EGCG), a major bioactive ingredient of green tea, is well-known to alleviate inflammation. Whether EGCG confers protection against SARA-induced inflammation and the underlying mechanisms are unknown.ResultsIn vivo, eight ruminally cannulated Holstein cows in mid-lactation were randomly assigned to either a low-concentrate (40%) diet (CON) or a high-concentrate (60%) diet (HC) for 3 weeks to induce SARA (n = 4). Cows with SARA had greater serum concentrations of tumor necrosis factor (TNF)-α and interleukin-6, and epithelium had histological signs of damage. In vitro, immortalized bovine ruminal epithelial cells (BREC) were treated with lipopolysaccharide (LPS) to imitate the inflammatory damage caused by SARA. Our data revealed that BREC treated with 10 µg/mL LPS for 6 h successfully induce a robust inflammatory response as indicated by increased phosphorylation of IκBα and nuclear factor kappa-B (NF-κB) p65. Pre-treatment of BREC with 50 µmol/L EGCG for 6 h before LPS challenge promoted the degradation of NLR family pyrin domain containing 3 (NLRP3) inflammasome through activation of autophagy, which further repressed activation of NF-κB pathway targeting Toll-like receptor 4 (TLR4). Analyses also revealed that the ECGG upregulated tight junction (TJ) protein expression upon incubation with LPS.ConclusionsSubacute ruminal acidosis causes ruminal epithelium injury and systemic inflammation in dairy cows. However, the anti-inflammatory effects of EGCG help preserve the integrity of the epithelial barrier through activating autophagy when BREC are exposed to LPS. Thus, EGCG could potentially serve as an effective therapeutic agent for SARA-associated inflammation.Graphical

Post-milking application of a Lacticaseibacillus paracasei strain impacts bovine teat microbiota while preserving the mammary gland physiology and immunity.

Bovine mastitis (BM) is a major disease in dairy industry. The current approaches- mainly antibiotic treatments- are not entirely effective and may contribute to antimicrobial resistance dissemination, rising the need for alternative treatment. The present study aims to evaluate the impact of post-milking application of Lacticaseibacillus paracasei CIRM BIA 1542 (Lp1542) on the teat skin (TS) of 20 Holstein cows in mid lactation, in order to reinforce the barrier effect of the microbiota naturally present on the teat. Treatment (Lp1542, iodine or no treatment) was applied post-milking twice a day on the 4 teats of healthy animals for 15 days. Blood and milk samples, and TS swabs were collected at day (D)1, D8, D15 and D26 before morning milking and at D15 before evening milking (D15E) to evaluate Lp1542 impact at the microbial, immune and physiological levels. Lp1542 treatment resulted in a higher lactic acid bacteria and total microbial populations on TS and in foremilk (FM) at D15(E) compared with iodine treatment. Metabarcoding analysis revealed changes in the composition of TS and FM microbiota, beyond a higher Lacticaseibacillus abundance. This included a higher abundance of Actinobacteriota, including Bifidobacterium, and a lower abundance of Pseudomonadota on TS of Lp1542 compared with iodine-treated quarters. In addition, Lp1542 treatment did not trigger any major inflammatory response in the mammary gland, except interleukin 8 production and expression which tended to be slightly higher in Lp1542-treated cows compared with the others. Finally, Lp1542 treatment had no impact on the mammary epithelium functionality (milk yield and composition) and integrity (epithelial cell exfoliation into milk and milk Na+/K+ ratio). Altogether, these results indicate that a topical treatment with Lp1542 is safe with regard to mammary gland physiology and immune system, while impacting its microbiota, inviting us to further explore its effectiveness for mastitis prevention.

The Effect of a Direct Fed Microbial on Liveweight and Milk Production in Dairy Cattle.

This longitudinal study aimed to quantify the effects of dietary supplementation of a direct-fed microbial (DFM) consisting of three lactobacilli isolates on milk yield, milk fat and protein yields, somatic cell count (SCC), and liveweight in a single dairy herd in Australia. A total of 150 dairy cows were randomly selected based on parity and days in milk and divided into two groups: control (n = 75) and DFM treatment (n = 75). Throughout the study, the two groups of cows were housed separately in a dry lot yard, and each group had their own feeding area. For the DFM treatment group, selected cows in mid-lactation were supplemented with 10 mL/cow/day of the DFM via top dressing of the feed for the remainder of the lactation and through the dry period, extending into subsequent lactation. The control group had no supplementation. The milk yield and liveweight were recorded daily. Milk samples were collected every two months for milk component analysis (fat, protein, and somatic cell count [SCC]). The DFM-treated cows gained more liveweight across the study (19.40 kg, 95% CI 0.44 kg; 38.30 kg, p = 0.05) compared to the control cows. In the second production year, the DFM-treated cows mobilized more liveweight (-6.06 kg, 95% CI -10.49 kg; -1.61 kg, p = 0.01) and produced more milk (0.39 L/d 95% CI 0.10; 0.89, p = 0.05). Over a full lactation, DFM cows yielded at least 258 L (95% CI 252 L; 265 L) more milk than controls. No significant differences were found in fat and protein yield or SCC. This study suggests that consistent and ongoing supplementation with a Lacticaseibacillus- and Lentilactobacillus-based DFM could have a positive effect on milk production, but further research is needed to understand the underlying mechanism.

Intravenous lipopolysaccharide challenge in early- versus mid-lactation dairy cattle. II: The production and metabolic responses

Most immunometabolic research uses mid-lactation (ML) cows. Cows in early lactation (EL) are in a presumed state of immune suppression/dysregulation and less is known about how they respond to a pathogen. Study objectives were to compare the production and metabolic responses to i.v. LPS and to differentiate between the direct effects of immune activation and the indirect effects of illness-induced hypophagia in EL and ML cows. Cows in EL (n = 11; 20 ± 2 DIM) and ML (n = 12; 131 ± 31 DIM) were enrolled in a 2 × 2 factorial design containing 2 experimental periods (P). During P1 (3 d), cows were fed ad libitum and baseline data were collected. At the initiation of P2 (3 d), cows were randomly assigned to 1 of 2 treatments by lactation stage (LS): (1) EL (EL-LPS; n = 6) or ML (ML-LPS; n = 6) cows administered i.v. a single bolus of 0.09 µg LPS/kg of BW; Escherichia coli O55:B5 or (2) pair-fed (PF) EL (EL-PF; n = 5) or ML (ML-PF; n = 6) cows administered i.v. saline. Administering LPS decreased DMI and this was more severe in EL-LPS than ML-LPS cows (34% and 11% relative to baseline, respectively). By design, P2 DMI patterns were similar in the PF groups compared with their LPS counterparts. Milk yield decreased following LPS (42% on d 1 relative to P1) and despite an exacerbated decrease in EL-LPS cows on d 1 (25% relative to ML-LPS), remained similar between LS from d 2 to 3. The EL-LPS cows had increased milk fat content, but no difference in protein and lactose percentages compared with ML-LPS cows. Further, cumulative ECM yield was increased (21%) in EL-LPS compared with ML-LPS cows. During P2, EL-LPS cows had a more intense increase in MUN and BUN than ML-LPS and EL-PF cows. Administering LPS did not cause hypoglycemia in either EL-LPS or ML-LPS cows, but glucose was increased (33%) in EL-LPS compared with EL-PF. Hyperinsulinemia occurred after LPS, and insulin was further increased in ML-LPS than EL-LPS cows (2.2-fold at 12 h peak). During P2, circulating glucagon increased only in EL-LPS cows (64% relative to all other groups). Both EL groups had increased NEFA at 3 and 6 h after LPS from baseline (56%), but NEFA in EL-LPS cows gradually returned to baseline thereafter and were reduced relative to EL-PF until 36 h (50% from 12 to 24 h). Alterations in BHB did not differ between ML groups, but EL-LPS had reduced BHB compared with EL-PF from 24 to 72 h (51%). Results indicate that there are distinct LS differences in the anorexic and metabolic responses to immune activation. Collectively, EL cows are more sensitive to the catabolic effects of LPS than ML cows, but these exacerbated metabolic responses appear coordinated to fuel an augmented immune system while simultaneously supporting milk synthesis.

Intramammary lipopolysaccharide challenge in early- versus mid-lactation dairy cattle: Immune, production, and metabolic responses

Study objectives were to compare the immune response, metabolism and production following intramammary lipopolysaccharide (IMM LPS) administration in early and mid-lactation cows. Early (E-LPS; n = 11; 20 ± 4 d in milk [DIM]) and mid- (M-LPS; n = 10; 155 ± 40 DIM) lactation cows were enrolled in an experiment consisting of 2 periods (P). During P1 (5 d) cows were fed ad libitum and baseline data were collected, including liver and muscle biopsies. At the beginning of P2 (3 d) cows received 10 mL sterile saline containing 10 µg of LPS from Escherichia coli O111:B4/mL into the left rear quarter of the mammary gland, and liver and muscle biopsies were collected at 12 h post-LPS. Tissues were analyzed for metabolic flexibility, which measures substrate switching capacity from pyruvic acid to palmitic acid oxidation. Data were analyzed with the MIXED procedure in SAS 9.4. Rectal temperature was assessed hourly for the first 12 h post-LPS and every 6 h thereafter for the remainder of P2. All cows developed a febrile response following LPS, but E-LPS had a more intense fever than M-LPS cows (0.7°C at 5 h after LPS). Blood samples were collected at 0, 3, 6, 9, 12, 24, 36, 48, and 72 h post-LPS for analysis of systemic inflammation and metabolism parameters. Total serum Ca decreased after LPS (26% at 6 h nadir) but did not differ by lactation stage (LS). Circulating neutrophils decreased, then increased post-LPS in both LS, but E-LPS had exaggerated neutrophilia (56% from 12 to 48 h) compared with M-LPS. Haptoglobin increased after LPS (15-fold) but did not differ by LS. Many circulating cytokines were increased post-LPS, and IL-6, IL-10, TNF-α, MCP-1, and IP-10 were further augmented in E-LPS compared with M-LPS cows. Relative to P1, all cows had reduced milk yield (26%) and dry matter intake (DMI; 14%) on d 1 that did not differ by lactation stage (LS). Somatic cell score increased rapidly in response to LPS regardless of LS and gradually decreased from 18 h onwards. Milk component yields decreased after LPS. However, E-LPS had increased fat (11%) and tended to have increased lactose (8%) yield compared with M-LPS cows throughout P2. Circulating glucose was not affected by LPS. Nonesterified fatty acids (NEFA) decreased in E-LPS (29%) but not M-LPS cows. β-hydroxybutyrate (BHB) slightly increased (14%) over time post-LPS regardless of LS. Insulin increased after LPS in all cows, but E-LPS had blunted hyperinsulinemia (52%) compared with M-LPS cows. Blood urea nitrogen (BUN) increased after LPS and the relative change in BUN was elevated in E-LPS cows compared with M-LPS cows (36 and 13%, respectively, from 9 to 24 h). During P1, metabolic flexibility was increased in liver and muscle in early lactating cows compared with mid-lactation cows, but 12 h post-LPS, metabolic flexibility was reduced and did not differ by LS. In conclusion, IMM LPS caused severe immune activation and E-LPS cows had a more intense inflammatory response compared with M-LPS cows, but the effects on milk synthesis was similar between LS. Some parameters of the E-LPS metabolic profile suggest continuation of metabolic adjustments associated with early lactation to support both a robust immune system and milk synthesis.

Intravenous lipopolysaccharide challenge in early- versus mid-lactation dairy cattle. I: The immune and inflammatory responses

Cows in early lactation (EL) are purportedly immune suppressed, which renders them more susceptible to disease. Thus, the study objective was to compare key biomarkers of immune activation from i.v. LPS between EL and mid-lactation (ML) cows. Multiparous EL (20 ± 2 DIM; n = 11) and ML (131 ± 31 DIM; n = 12) cows were enrolled in a 2 × 2 factorial design and assigned to 1 of 2 treatments by lactation stage (LS): (1) EL (EL-LPS; n = 6) or ML (ML-LPS; n = 6) cows administered a single LPS bolus from Escherichia coli O55:B5 (0.09 µg/kg of BW), or (2) pair-fed (PF) EL (EL-PF; n = 5) or ML (ML-PF; n = 6) cows administered i.v. saline. After LPS administration, cows were intensely evaluated for 3 d to analyze their response and recovery to LPS. Rectal temperature increased in LPS relative to PF cows (1.1°C in the first 9 h), and the response was more severe in EL-LPS relative to ML-LPS cows (2.3 vs. 1.3°C increase at 4 h post-LPS; respectively). Respiration rate increased only in EL-LPS cows (47% relative to ML-LPS in the first hour post-LPS). Circulating tumor necrosis factor-α, IL-6, monocyte chemoattractant protein-1, macrophage inflammatory protein (MIP)-1α, MIP-1β, and IFN-γ-inducible protein-10 increased within the first 6 h after LPS and these changes were exacerbated in EL-LPS relative to ML-LPS cows (6.3-fold, 4.8-fold, 57%, 93%, 10%, and 61%, respectively). All cows administered LPS had decreased circulating iCa relative to PF cows (34% at the 6 h nadir), but the hypocalcemia was more severe in EL-LPS than ML-LPS cows (14% at 6 h nadir). In response to LPS, neutrophils decreased regardless of LS, then increased into neutrophilia by 24 h in all LPS relative to PF cows (2-fold); however, the neutrophilic phase was augmented in EL- compared with ML-LPS cows (63% from 24 to 72 h). Lymphocytes and monocytes rapidly decreased then gradually returned to baseline in LPS cows regardless of LS; however, monocytes were increased (57%) at 72 h in EL-LPS relative to ML-LPS cows. Platelets were reduced (46%) in LPS relative to PF cows throughout the 3-d following LPS, and from 24 to 48 h, platelets were further decreased (41%) in EL-LPS compared with ML-LPS. During the 3-d following LPS, serum amyloid A (SAA), LPS-binding protein (LBP), and haptoglobin (Hp) increased in LPS compared with PF groups (9-fold, 72%, and 153-fold, respectively), and the LBP and Hp responses were more exaggerated in EL-LPS than ML-LPS cows (85 and 79%, respectively) whereas the SAA response did not differ by LS. Thus, our data indicates that EL immune function does not appear "suppressed," and in fact many aspects of the immune response are seemingly functionally robust.

Influence of Heat Stress on Body Surface Temperature and Blood Metabolic, Endocrine, and Inflammatory Parameters and Their Correlation in Cows.

This study aimed to determine whether heat stress affected the values and correlations of metabolic, endocrinological, and inflammatory parameters as well as the rectal and body surface temperature of cows in the early and middle stages of lactation. This experiment was conducted in May (thermoneutral period), June (mild heat stress), and July (moderate to severe heat stress). In each period we included 15 cows in early lactation and 15 in mid-lactation. The increase in rectal and body surface temperatures (°C) in moderate to severe heat stress compared to the thermoneutral period in different regions was significant (p < 0.01) and the results are presented as mean and [95%CI]: rectal + 0.9 [0.81-1.02], eye + 6 [5.74-6.25], ear + 13 [11.9-14.0], nose + 3.5 [3.22-3.71], forehead + 6.6 [6.43-6.75], whole head + 7.5 [7.36-7.68], abdomen + 8.5 [8.25-8.77], udder + 7.5 [7.38-7.65], front limb + 6 [5.89-6.12], hind limb + 3.6 [3.46-3.72], and whole body + 9 [8.80-9.21]. During heat stress (in both mild and moderate to severe stress compared to a thermoneutral period), an increase in the values of extracellular heat shock protein 70 (eHsp70), tumor necrosis factor α (TNFα), cortisol (CORT), insulin (INS), revised quantitative insulin sensitivity check index (RQUICKI), urea, creatinine, total bilirubin, aspartate transpaminase (AST), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), and creatin kinase (CK) occurred, as well as a decrease in the values of triiodothyronine (T3), thyroxine (T4), non-esterified fatty acids (NEFA), glucose (GLU), β-Hydroxybutyrate (BHB), calcium, phosphorus, total protein (TPROT), albumin (ALB), triglycerides (TGCs), and cholesterol (CHOL). In cows in early lactation compared to cows in mid-lactation, there was a significantly larger increase (p < 0.01) in the values of eHsp70, TNFα, GLU, RQUICKI, and GGT, while the INS increase was smaller during the three experimental periods. The decrease in the values of Ca, CHOL, and TGC was more pronounced in cows in early lactation compared to cows in mid-lactation during the three experimental periods. Rectal temperature was related to eHsp70 (r = 0.38, p < 0.001) and TNFα (r = 0.36, p < 0.01) and showed non-significant poor correlations with other blood parameters. Blood parameters correlate with body surface temperature, with the following most common results: eHsp70 and TNFα showed a moderately to strongly significant positive correlation (r = 0.79-0.96, p < 0.001); CORT, INS, and Creat showed fairly to moderately significant positive correlations; T3, T4, NEFA and GLU showed fairly to moderately significant negative correlations (r = 0.3-0.79; p < 0.01); RQUICKI, urea, AST, and GGT showed fairly and significantly positive correlations; and TGC, CHOL, TPROT, and ALB showed fairly and significantly negative correlations (r = 0.3-0.59; p < 0.01). Measuring the surface temperature of the whole body or head can be a useful tool in evaluating the metabolic response of cows because it has demonstrated an association with inflammation (TNFα, eHsp70), endocrine response (CORT, T3, T4), the increased use of glucose and decreased use of lipids for energy purposes (INS, NEFA, GLU, and RQUICKI), and protein catabolism (ALB, TPROT, urea, Creat), which underlies thermolysis and thermogenesis in cows under heat stress. In future research, it is necessary to examine the causality between body surface area and metabolic parameters.

Effects of pellet starch levels in automatic milking systems on rumen fermentation, plasma metabolites, and milk production in mid-lactation cows.

The aim of this study was to evaluate whether the starch levels in pellets fed to cows in automatic milking systems (AMS) affect subacute ruminal acidosis (SARA) occurrence and metabolite parameters. Twenty-four lactating cows (124.4 ± 49.9 days in milk) were studied in a crossover design with two periods of 21 days each and two treatment groups-a control group fed AMS pellets containing 30.0% of starch dry matter (DM) and an experimental group fed AMS pellets containing 23.5% of starch DM. All cows received the same partial mixed ration (PMR). The 1-hr mean ruminal pH in both groups decreased over 4 hr after feeding on PMR but recovered by the next morning. The ruminal pH was unaffected by either treatment, and both groups developed SARA. The groups had no significant differences in the concentrations of ruminal volatile fatty acids, lipopolysaccharides, plasma acute-phase proteins, other metabolites, and hormones. The milk yield and composition were not different in both groups. Feeding low-starch pellets in the AMS did not contribute to the risk of SARA occurrence in cows and had no additive effects on rumen fermentation, plasma metabolites, or milk production.

Effects of heat stress on feed intake, milk yield, milk composition, and feed efficiency in dairy cows: A meta-analysis

Heat stress compromises dairy production by decreasing feed intake and milk yield, and it may also alter milk composition and feed efficiency. However, little information is available for evaluating such effects across different levels of heat stress and cows enrolled in heat stress studies. The objectives of this study were to evaluate the effects of heat stress on dry matter intake (DMI), energy-corrected milk (ECM), milk composition, and feed efficiency (kg ECM/kg DMI) and to investigate the relationship between such effects and heat stress intervention and animal characteristics by using meta-analytical approaches. Data from 31 studies (34 trials) fulfilled the inclusion criteria and were used for analysis. Results showed that heat stress decreased DMI, ECM, and milk protein concentration, but did not alter milk fat concentration or feed efficiency. Meta-regression confirmed that such reductions in DMI and ECM were significantly associated with increasing temperature-humidity index (THI). Over the period of heat stress, for each unit increase in THI, DMI and ECM decreased by 4.13% and 3.25%, respectively, in mid-lactation cows. Regression models further revealed the existence of a strong interaction between THI and lactation stage, which partially explained the large heterogeneity in effect sizes of DMI and ECM. The results indicated a need for more research on the relationship between the effect of heat stress and animal characteristics. This study calls for the implementation of mitigation strategies in heat-stressed herds due to the substantial decrease in productivity.

Interaction between drinker density and cow social dominance affects drinking behavior

Dairy cow welfare and milk production levels can be negatively affected when animals lack access to sufficient amounts of good-quality drinking water. Farms vary greatly in how well they meet this standard. Here, we explored how the interaction between drinker density and social dominance level influenced the drinking behaviors of a group of 40 mid-lactation cows. We recorded cow drinking behavior and competitive outcomes (i.e., number of replacements) at the drinkers during the final 5 d of 2 drinker density treatments, applied successively (12 drinkers, then 4 drinkers available). Four social dominance categories were defined by applying the quartile classification method to the normalized David's score obtained from the low-density treatment data (NormDS values). The high- and low-density treatments used approximately 56 (±16) versus 119 (±31) replacements per drinker per day. Subordinate cows were lighter and younger than dominant cows. Dominant cows drank around 5 L more per day in the low- versus high-density treatment. Furthermore, both drinking rate and daily drinking frequency were higher in the low-density treatment, with the most pronounced differences seen for subordinate cows. In the low-density treatment, mid-subordinate cows shifted their drinking times, visiting the drinkers 1 to 2 h after peak water consumption by the group. The results for the low-density treatment thus indicate that the cows were experiencing more intense competition for water, leading to behavioral responses that were related to social dominance. Our findings underscore the need to more broadly explore the effects of increasing drinker numbers under a variety of rearing conditions to provide farmers with better technical recommendations.

73 Effects of Fatty Acids Sources and Lysophospholipids on Nutrient Digestibility in Lactating Cows

Abstract Fatty acids (FA) are a common supplement for lactating cows to increase energy supply, potentially improving feed efficiency and milk and milk fat yields. However, different FA can cause varying responses probably because of different FA digestibility and interactions with other nutrients (e.g., fibers). Lysophospholipids (LPL) are natural emulsifiers and LPL as a feed additive have shown increases in FA digestibility and nutrient absorption in non-ruminant animals. This study aimed to investigate the effects of FA sources with LPL supplementation on nutrient digestibility in lactating cows. Mid-lactation cows (n = 48) were allocated into 12 blocks, and cows in each block were randomly assigned to the following dietary treatments:1) a diet with a FA supplement [1.8% of dietary dry matter (DM)] mainly containing palmitic acid (PA); 2) the PA diet with LPL (0.003% of dietary DM); 3) a diet with a FA supplement (1.8% of dietary DM) mainly containing stearic acid (SA); and 4) the SA diet with LPL (0.003% of DM). The experiment was conducted for 6 weeks, and spot samples of feces and urine were collected over 3 days (every 3 hours within a 24-hour cycle) during the last week of the experiment to determine nutrient digestibility and utilization. Data were analyzed using the mixed model with FA source, LPL, and their interaction as fixed effects and block as a random effect. The digestibility of DM, organic matter, and crude protein were not affected by the FA source and no interaction between FA and LPL was observed. However, PA had greater neutral detergent fiber (NDF) digestibility (41.4 vs. 38.8%; P = 0.04) than SA with no interaction between the FA source and LPL. In addition, as expected, PA decreased (P &amp;lt; 0.01) the digestibility of 16-carbon FA compared with SA due to the greater C16:0 supply, and SA decreased (P &amp;lt; 0.01) that of 18-carbon FA compared with PA due to the greater C18:0 supply. Supplementation of LPL did not affect digestibility of DM, OM, CP, and NDF, but increased total FA digestibility (64.7 vs. 60.5%; P = 0.02) compared with no LPL. In addition, LPL increased 18-carbon FA digestibility (68.2 vs. 74.1%; P &amp;lt; 0.01) compared with no LPL. An interaction between LPL and FA was observed for 16-carbon digestibility because LPL increased 16-carbon digestibility with SA but not with PA. In conclusion, PA supplementation improved fiber digestibility compared with SA, which could be a factor for increased productivity (e.g., milk fat). Supplementation of LPL was effective in increasing FA digestibility and may have potential to increase dietary energy supply or provide a similar energy supply with less dietary fat content in practice. However, more studies are needed to understand different interactions of LPL with FA sources.

Serum metabolome differences associated with subclinical intramammary infection caused by Streptococcus agalactiae and Prototheca spp. in multiparous dairy cows

Mastitis is one of the most significant diseases in dairy cows and causes several economic losses. Somatic cell count (SCC) is often used as an indirect diagnostic tool for mastitis, especially for subclinical mastitis (SCM) where no symptoms or signs can be detected. Streptococcus agalactiae is one of the main causes of contagious mastitis, while Prototheca spp. is an alga inducing environmental mastitis that is not always correlated with increased milk SCC. The aim of this study was to evaluate the changes in the metabolomic profile of blood in relation to subclinical intramammary infection (sIMI) in dairy cows. In addition, differences due to the etiologic agent causing mastitis were also considered. Forty Holstein-Friesian dairy cows in mid and late lactation were enrolled in this study with a cross-sectional design. Based on the bacteriological examination of milk, the animals were divided into 3 groups: Group CTR (control group; n = 16); Group A (affected by SCM with IMI of Streptococcus agalactiae; n = 17); and Group P (affected by SCM with IMI of Prototheca spp.; n = 7). Blood samples were collected in tubes containing clot activator from jugular vein. The serum aliquot was stored until metabolomic analysis by 1H-NMR. Statistical analysis was conducted fitting a linear model with the group as fixed effect and SCC as covariate. Forty-two metabolites were identified and among them, 10 were significantly different among groups. Group A and P showed greater level of His and lactose, and lower level of acetate, Asn, and dimethylamine compared with Group CTR. Group A showed high level of Val, while the Group P showed also high level of Cit and methylguanidine, and lower level of 3-hydroxybutyrate, acetone, allantoin, carnitine, citrate, and ethanol. These metabolites were related to ruminal fermentations, energy metabolism, urea synthesis and metabolism, immune and inflammatory response, and mammary gland permeability. These results are suggestive of a systemic involvement of sIMI and that the metabolic profile of animals with SCM undergoes changes related to the etiological agent of mastitis.

Effects of oyster glycogen intramammary challenge on primiparous cow milk somatic cell counts, milk yields, and milk composition

Mastitis is a common and costly disease in the dairy industry that reduces milk production in affected mammary glands. The local mechanisms that result in reduced milk production of affected mammary glands are incompletely understood; elucidation of these mechanisms is dependent on the use of hypothesis testing studies, but few experimental models exist. The objective of this study was to develop a mastitis challenge model, using a split udder design, to reduce milk yields by approximately 15% in udder halves challenged with oyster glycogen, a known inducer of leukocyte recruitment, relative to udder halves treated with saline. Four primiparous Holstein cows in mid lactation were used. One udder half of each cow was randomly selected and challenged with oyster glycogen (OYGLN), and the opposite udder half was treated with saline (SAL). Milk yields and components were measured at each milking (3×/d) for 3 d postchallenge. No signs of clinical mastitis were observed. Milk somatic cell scores, yields, and components were similar between OYGLN and SAL udder halves at time of challenge. Milk somatic cell scores markedly increased in OYGLN halves postchallenge and were greater than SAL halves for the duration of the trial. Lactose concentrations of OYGLN udder halves were transiently lower than in SAL udder halves, but protein concentrations were greater at 2 milkings postchallenge in OYGLN halves. Milk yields and energy-corrected milk yields did not differ between OYGLN and SAL udder halves overall, nor at any postchallenge milking. A single intramammary challenge of oyster glycogen was unsuccessful in eliciting a disparity in milk yields between challenged and saline control udder halves despite the marked leukocyte infiltration observed in the former. These results indicate an incomplete understanding of how milk yields are reduced in mammary glands affected by subclinical mastitis and that transient somatic cell recruitment and infiltration alone do not directly reduce milk yields during subclinical mastitis.