Objectives were to evaluate effects of supplemental zinc hydroxychloride (HYD; Micronutrients, Indianapolis, IN) on gut permeability, metabolism, and inflammation during feed restriction (FR). Holstein cows (n = 24; 159 ± 8 d in milk; parity 3 ± 0.2) were enrolled in a 2 × 2 factorial design and randomly assigned to 1 of 4 treatments: (1) ad libitum fed (AL) and control diet (ALCON; 75 mg/kg Zn from zinc sulfate; n = 6); (2) ad libitum fed and HYD diet (ALHYD; 75 mg/kg Zn from HYD; n = 6); (3) 40% of ad libitum feed intake and control diet (FRCON; n = 6); or (4) 40% of ad libitum feed intake and HYD diet (FRHYD; n = 6). Prior to study initiation, cows were fed their respective diets for 21 d. The trial consisted of 2 experimental periods (P) during which cows continued to receive their respective dietary treatments. Period 1 (5 d) served as the baseline for P2 (5 d), during which cows were fed ad libitum or restricted to 40% of P1 feed intake. In vivo total-tract permeability was evaluated on d 4 of P1 and on d 2 and 5 of P2, using the paracellular permeability marker chromium (Cr)-EDTA. All cows were euthanized at the end of P2 to assess intestinal architecture. As anticipated, FR cows lost body weight (~46 kg), entered into calculated negative energy balance (-13.86 Mcal/d), and had decreased milk yield. Circulating glucose, insulin, and glucagon decreased, and nonesterified fatty acids and β-hydroxybutyrate increased in FR relative to AL cows. Relative to AL cows, FR increased lipopolysaccharide-binding protein, serum amyloid A (SAA), and haptoglobin (Hp) concentrations (2-, 4-, and 17-fold, respectively); and peak SAA and Hp concentrations were observed on d 5. Circulating SAA and Hp from FRHYD tended to be decreased (47 and 61%, respectively) on d 5 relative to FRCON. Plasma Cr area under the curve increased (32%) in FR treatments on d 2 and tended to be increased (17%) on d 5 of P2 relative to AL treatments. No effects of diet were observed on Cr appearance. Relative to AL cows, FR increased jejunum villus width and decreased jejunum crypt depth and ileum villus height and crypt depth. Relative to FRCON, ileum villus height tended to increase in FRHYD cows. Feed restriction tended to decrease jejunum and ileum mucosal surface area, but the decrease in the ileum was ameliorated by dietary HYD. In summary, FR induced gut hyperpermeability to Cr-EDTA, and feeding HYD appeared to benefit some key metrics of barrier integrity.

Infection and inflammation impede efficient animal productivity. The activated immune system ostensibly requires large amounts of energy and nutrients otherwise destined for synthesis of agriculturally relevant products. Accurately determining the immune system’s in vivo energy needs is difficult, but a better understanding may facilitate developing nutritional strategies to maximize productivity. The study objective was to estimate immune system glucose requirements following an i.v. lipopolysaccharide (LPS) challenge. Holstein steers (148 ± 9 kg; n = 15) were jugular catheterized bilaterally and assigned to 1 of 3 i.v. treatments: control (CON; 3 mL saline; n = 5), LPS-administered controls (LPS-C; E. coli 055:B5; 1.5 mg/kg BW; n = 5), and LPS + euglycemic clamp (LPS-Eu; 1.5 mg/kg BW; 50% dextrose infusion to maintain euglycemia; n = 5). In LPS-Eu steers, postbolus blood samples were analyzed for glucose every 10 min. Dextrose infusion rates were adjusted to maintain euglycemia for 720 min. All steers were fasted during the challenge. Samples for later analysis were obtained at 180, 360, 540, and 720 min relative to LPS administration. Rectal temperature was increased ∼0.5°C in both LPS treatments relative to CON steers (P = 0.01). Steers in both LPS treatments were hyperglycemic for ∼3 h postbolus; thereafter, blood glucose was markedly decreased (30%; P < 0.01) in LPS-C relative to both CON and LPS-Eu steers. A total of 516 ± 65 g of infused glucose was required to maintain continuous euglycemia in LPS-Eu steers. Circulating insulin increased in LPS-C and LPS-Eu steers relative to CON (∼70% and ∼20 fold, respectively; P < 0.01). Circulating NEFA increased similarly with time for both CON and LPS-C compared to LPS-Eu steers (∼43%; P < 0.01). Plasma L-lactate and LPS binding protein increased (∼198 and ∼90%, respectively; P < 0.01) and ionized calcium decreased (18%; P < 0.01) in both LPS treatments relative to CON steers. Circulating white blood cells decreased initially in LPS-Eu and LPS-C relative to controls (180 min; 85%) followed by a progressive increase with time (P = 0.02). Blood neutrophils followed the same pattern; however, at 720 min, neutrophils were decreased in LPS-Eu compared to LPS-C, resulting in a decreased neutrophil-to-lymphocyte ratio (54%; P = 0.03). The large amount of glucose needed to maintain euglycemia indicates extensive repartitioning of nutrients away from growth and the importance of glucose as a fuel for the immune system. Copyright © 2016. American Society of Animal Science .

Postruminal intestinal barrier dysfunction caused by excessive hindgut fermentation may be a source of peripheral inflammation in dairy cattle. Therefore, the study objectives were to evaluate the effects of isolated hindgut acidosis on metabolism, inflammation, and production in lactating dairy cows. Five rumen-cannulated lactating Holstein cows (32.6 ± 7.2 kg/d of milk yield, 242 ± 108 d in milk; 642 ± 99 kg of body weight; 1.8 ± 1.0 parity) were enrolled in a study with 2 experimental periods (P). During P1 (4 d), cows were fed ad libitum a standard lactating cow diet (26% starch dry matter) and baseline data were collected. During P2 (7 d), all cows were fed the same diet ad libitum and abomasally infused with 4 kg/d of pure corn starch (1 kg of corn starch + 1.25 L of H₂O/infusion at 0600, 1200, 1800, and 0000 h). Effects of time (hour relative to the first infusion or day) relative to P1 were evaluated using PROC MIXED in SAS (version 9.4; SAS Institute Inc.). Infusing starch markedly reduced fecal pH (5.84 vs. 6.76) and increased fecal starch (2.2 to 9.6% of dry matter) relative to baseline. During P2, milk yield, milk components, energy-corrected milk yield, and voluntary dry matter intake remained unchanged. At 14 h, plasma insulin and β-hydroxybutyrate increased (2.4-fold and 53%, respectively), whereas circulating glucose concentrations remained unaltered. Furthermore, blood urea nitrogen increased at 2 h (23%) before promptly decreasing below baseline at 14 h (13%). Nonesterified fatty acids tended to decrease from 2 to 26 h (40%). Circulating white blood cells and neutrophils increased on d 4 (36 and 73%, respectively) and somatic cell count increased on d 5 (4.8-fold). However, circulating serum amyloid A and lipopolysaccharide-binding protein concentrations were unaffected by starch infusions. Despite minor changes in postabsorptive energetics and leukocyte dynamics, abomasal starch infusions and the subsequent hindgut acidosis had little or no meaningful effects on biomarkers of immune activation or production variables.

Heat-stress-induced inflammation may be ameliorated by antioxidant supplementation due to the purported effects of increased production of reactive oxygen species or oxidative stress on the gastrointestinal tract barrier. Thus, study objectives were to evaluate whether antioxidant supplementation [AGRADO Plus 2.0 (AP); EW Nutrition] affects metabolism and inflammatory biomarkers in heat-stressed lactating dairy cows. Thirty-two mid-lactation multiparous Holstein cows were assigned to 1 of 4 dietary-environmental treatments: (1) thermoneutral (TN) conditions and fed a control diet (TN-CON; n = 8), (2) TN and fed a diet with AP (10 g antioxidant; n = 8), (3) heat stress (HS) and fed a control diet (HS-CON; n = 8), or (4) HS and fed a diet with AP (HS-AP; n = 8). The trial consisted of a 23-d prefeeding phase and 2 experimental periods (P). Respective dietary treatments were top-dressed starting on d 1 of the prefeeding period and continued daily throughout the duration of the experiment. During P1 (4 d), baseline data were collected. During P2 (7 d), HS was artificially induced using an electric heat blanket (Thermotex Therapy Systems Ltd.). During P2, the effects of treatment, day, and treatment-by-day interaction were assessed using PROC MIXED of SAS (SAS Institute Inc.). Heat stress (treatments 3 and 4) increased rectal, vaginal, and skin temperatures (1.2°C, 1.1°C, and 2.0°C, respectively) and respiration rate (33 breaths per minute) relative to TN cows. As expected, HS decreased dry matter intake, milk yield, and energy-corrected milk yield (32%, 28%, and 28% from d 4 to 7, respectively) relative to TN. There were no effects of AP on body temperature indices or production. Milk fat, protein, and lactose concentrations remained unaltered by HS or AP; however, milk urea nitrogen was increased during HS regardless of AP supplementation (26% relative to TN). Circulating glucose remained unchanged by HS, AP, or time. Additionally, HS decreased circulating glucagon (29% from d 3 to 7 relative to TN), but there was no additional effect of AP. There was a tendency for nonesterified fatty acid concentrations to be increased in HS-AP cows throughout P2 (60% relative to TN-CON), whereas it remained similar in all other treatments. Blood urea nitrogen increased for both HS treatments from d 1 to 3 before steadily decreasing from d 5 to 7, with the overall increase being most pronounced in HS-CON cows (27% relative to TN-CON). Further, supplementing AP decreased blood urea nitrogen in HS-AP on d 3 relative to HS-CON (15%). Circulating serum amyloid A tended to be and lipopolysaccharide binding protein was increased by HS, but neither acute-phase protein was affected by AP. Overall, AP supplementation appeared to marginally alter metabolism but did not meaningfully alter inflammation during HS.

The objectives of this study were to evaluate the effects of replacing 40 mg/kg of Zn from Zn sulfate (control; CON) with Zn AA complex (AvZn) on metabolism and immunological responses following an intravenous lipopolysaccharide (LPS) challenge in lactating cows. Cows were randomly assigned to 1 of 4 treatments: (1) pair-fed (PF) control (PF-CON; 5 mL of saline; n = 5), (2) PF AvZn (PF-AvZn; 5 mL of saline; n = 5), (3) LPS euglycemic clamp control (LPS-CON; 0.375 μg of LPS/kg of BW; n = 5), and (4) LPS euglycemic clamp AvZn (LPS-AvZn; 0.375 μg of LPS/kg of BW; n = 5). Cows were enrolled in 3 experimental periods (P). During period 1 (3 d), cows received their respective dietary treatments and baseline data were obtained. During period 2 (P2; 2 d), a 12-h LPS euglycemic clamp was conducted or cows were PF to their respective dietary counterparts. During period 3 (P3; 3 d), cows received their dietary treatment and consumed feed ad libitum. Mild hyperthermia (1°C) was observed in LPS cows at 3 h postbolus. Throughout P2, the rectal temperature of LPS-AvZn cows was decreased (0.3°C) relative to LPS-CON cows. Administrating LPS decreased dry matter intake (47%) during P2, and by experimental design the pattern was similar in PF cohorts. During P3, dry matter intake from LPS cows remained decreased (15%) relative to PF cows. Milk yield from LPS cows decreased (54%) during P2 relative to PF cows, but it was similar during P3. During P2, somatic cell count increased 3-fold in LPS cows relative to PF controls. Dietary AvZn tended to decrease somatic cell count (70%) during P3 relative to LPS-CON cows. Insulin increased 7-fold in LPS cows at 12 h postbolus and remained increased (4-fold) for the duration of P2. Circulating glucagon from LPS cows increased (65%) during P2, and supplementing AvZn blunted the increase (30% relative to LPS-CON). During P2, circulating cortisol increased 7-fold post-LPS infusion relative to PF cows, and supplementing AvZn decreased cortisol (58%) from 6 to 48 h postbolus relative to LPS-CON cows. Administrating LPS increased circulating LPS-binding protein and serum amyloid A (3- and 9-fold, respectively) relative to PF cows. Compared with LPS-CON, LPS-AvZn cows had increased circulating serum amyloid A (38%) 24 h postbolus. The 12-h total glucose deficit was 36 and 1,606 g for the PF and LPS treatments, respectively, but was not influenced by Zn source. In summary, replacing a portion of the Zn sulfate with Zn AA complex appeared to reduce the inflammatory response but had no effect on the glucose deficit.

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.