Characterizing effects of feed restriction and glucagon-like peptide 2 administration on biomarkers of inflammation and intestinal morphology.

Inadequate feed consumption reduces intestinal barrier function in both ruminants and monogastrics. Objectives were to characterize how progressive feed restriction (FR) affects inflammation, metabolism, and intestinal morphology, and to investigate if glucagon-like peptide 2 (GLP2) administration influences the aforementioned responses. Twenty-eight Holstein cows (157 ± 9 d in milk) were enrolled in 2 experimental periods. Period 1 [5 d of ad libitum (AL) feed intake] served as baseline for period 2 (5 d), during which cows received 1 of 6 treatments: (1) 100% of AL feed intake (AL100; n = 3), (2) 80% of AL feed intake (n = 5), (3) 60% of AL feed intake (n = 5), (4) 40% of AL feed intake (AL40; n = 5), (5) 40% of AL feed intake + GLP2 administration (AL40G; 75 µg/kg of BW s.c. 2×/d; n = 5), or (6) 20% of AL feed intake (n = 5). As the magnitude of FR increased, body weight and milk yield decreased linearly. Blood urea nitrogen and insulin decreased, whereas nonesterified fatty acids and liver triglyceride content increased linearly with progressive FR. Circulating endotoxin, lipopolysaccharide binding protein, haptoglobin, serum amyloid A, and lymphocytes increased or tended to increase linearly with advancing FR. Circulating haptoglobin decreased (76%) and serum amyloid A tended to decrease (57%) in AL40G relative to AL40 cows. Cows in AL100, AL40, and AL40G treatments were euthanized to evaluate intestinal histology. Jejunum villus width, crypt depth, and goblet cell area, as well as ileum villus height, crypt depth, and goblet cell area, were reduced (36, 14, 52, 22, 28, and 25%, respectively) in AL40 cows compared with AL100 controls. Ileum cellular proliferation tended to be decreased (14%) in AL40 versus AL100 cows. Relative to AL40, AL40G cows had improved jejunum and ileum morphology, including increased villus height (46 and 51%), villus height to crypt depth ratio (38 and 35%), mucosal surface area (30 and 27%), cellular proliferation (43 and 36%), and goblet cell area (59 and 41%). Colon goblet cell area was also increased (48%) in AL40G relative to AL40 cows. In summary, progressive FR increased circulating markers of inflammation, which we speculate is due to increased intestinal permeability as demonstrated by changes in intestinal architecture. Furthermore, GLP2 improved intestinal morphology and ameliorated circulating markers of inflammation. Consequently, FR is a viable model to study consequences of intestinal barrier dysfunction and administering GLP2 appears to be an effective mitigation strategy to improve gut health.

Intentionally induced intestinal barrier dysfunction causes inflammation, affects metabolism, and reduces productivity in lactating Holstein cows.

Study objectives were to evaluate the effects of intentionally reduced intestinal barrier function on productivity, metabolism, and inflammatory indices in otherwise healthy dairy cows. Fourteen lactating Holstein cows (parity 2.6 ± 0.3; 117 ± 18 d in milk) were enrolled in 2 experimental periods. Period 1 (5 d) served as the baseline for period 2 (7 d), during which cows received 1 of 2 i.v. treatments twice per day: sterile saline or a gamma-secretase inhibitor (GSI; 1.5 mg/kg of body weight). Gamma-secretase inhibitors reduce intestinal barrier function by inhibiting crypt cell differentiation into absorptive enterocytes. During period 2, control cows receiving sterile saline were pair-fed (PF) to the GSI-treated cows, and all cows were killed at the end of period 2. Administering GSI increased goblet cell area 218, 70, and 28% in jejunum, ileum, and colon, respectively. In the jejunum, GSI-treated cows had increased crypt depth and reduced villus height, villus height-to-crypt depth ratio, cell proliferation, and mucosal surface area. Plasma lipopolysaccharide binding protein increased with time, and tended to be increased 42% in GSI-treated cows relative to PF controls on d 5 to 7. Circulating haptoglobin and serum amyloid A concentrations increased (585- and 4.4-fold, respectively) similarly in both treatments. Administering GSI progressively reduced dry matter intake (66%) and, by design, the pattern and magnitude of decreased nutrient intake was similar in PF controls. A similar progressive decrease (42%) in milk yield occurred in both treatments, but we observed no treatment effects on milk components. Cows treated with GSI tended to have increased plasma insulin (68%) and decreased circulating nonesterified fatty acids (29%) compared with PF cows. For both treatments, plasma glucose decreased with time while ß-hydroxybutyrate progressively increased. Liver triglycerides increased 221% from period 1 to sacrifice in both treatments. No differences were detected in liver weight, liver moisture, or body weight change. Intentionally compromising intestinal barrier function caused inflammation, altered metabolism, and markedly reduced feed intake and milk yield. Further, we demonstrated that progressive feed reduction appeared to cause leaky gut and inflammation.

Effect of monensin in lactating dairy cow diets at 2 starch concentrations.

The objective of this study was to determine the effects of monensin (M) supplementation on lactation performance of dairy cows fed diets of either reduced (RS) or normal (NS) starch concentrations as total mixed rations. One hundred twenty-eight Holstein and Holstein × Jersey cows (90 ± 33 d in milk) were stratified by breed and parity and randomly assigned to 16 pens of 8 cows each in a randomized controlled trial. Pens were then randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement of treatments. A 4-wk covariate adjustment period preceded the treatment period, with all pens receiving NS supplemented with 18 g of monensin/t of dry matter (DM). Following the 4-wk covariate adjustment period, cows were fed their assigned treatment diets of NS with M (18 g of monensin/t), NS with 0 g of monensin/t (C), RS with M, or RS with C for 12 wk. Actual starch concentrations for the RS and NS diets were 20.4 and 26.9% (DM basis), respectively. Mean dry matter intake (DMI; 27.0 kg/d) was unaffected by the treatments. Feeding M compared with C and NS compared with RS increased milk yield by 1.3 and 1.5 kg/d per cow, respectively. Milk protein percentage and yield and lactose yield were increased and milk urea nitrogen was decreased for NS compared with RS. Feeding M increased actual and component-corrected milk feed efficiencies (component-corrected milk yield/DMI) and lactose yield and tended to increase milk urea nitrogen compared with C. Milk protein percentage was decreased for M compared with C, but milk fat percentage and yield, protein yield, and lactose percentage were unaffected by M. We observed a tendency for a starch × monensin interaction for milk feed efficiency (actual milk yield/DMI); M tended to increase efficiency more for NS than for RS. Starch and monensin had minimal effects on milk fatty acid composition and yields. Feeding RS decreased milk and protein yields, but component-corrected milk yields and feed efficiencies were similar for RS and NS. Monensin increased feed efficiency and lactation performance for both dietary starch concentrations.

Effect of dietary fat blend enriched in oleic or linoleic acid and monensin supplementation on dairy cattle performance, milk fatty acid profiles, and milk fat depression.

The effect of feeding increasing levels of oleic and linoleic acid both independently and together, with or without monensin, on milk fat depression was evaluated. Fifty-six Holstein cows were blocked by parity and then were divided by milk production into 2 groups (high or low) of 14 cows each within each parity block. A cow pair of 1 high and 1 low production cow within each parity block was fed in a single electronic feeding gate. Gates (n = 28) were considered the experimental unit and were assigned to monensin (17.5 g/t of dry matter) or control as the main plot (n = 14 each). The 7 cow pairs in each of the fixed effect groups were further assigned to a sequence of fat blend diets as split plot. Seven fat blend treatments in the split plot 7 × 7 Latin square were no added fat (no fat) and diets with increasing levels of oleic or linoleic acid: low C18:1 + low C18:2 (LOLL); low C18:1 + medium C18:2 (LOML); low C18:1 + high C18:2 (LOHL); medium C18:1 + low C18:2 (MOLL); medium C18:1+medium C18:2 (MOML); and high C18:1+low C18:2 (HOLL). Monensin feeding did not affect milk yield or concentration and yield of milk fat. Feeding monensin decreased the proportion of C <16, increased the proportion of total C18, increased the proportion and yield of trans-10 C18:1, and increased the proportion of trans-10,cis-12 conjugated linoleic acid in milk fatty acids (FA). As dietary C18:1 or C18:2 increased beyond the concentration present in LOLL, milk fat concentration, milk fat yield, and proportion and yield of milk C <16 all decreased, and the proportion and yield of milk trans-10 C18:1 increased. A quadratic effect on milk fat concentration and yield was noticed for C18:2 feeding, but not for C18:1 feeding. When dietary contents of total FA and FA other than C18:1 and C18:2 were similar, C18:2-rich diets decreased milk fat concentration and yield compared with C18:1-rich diets (LOML vs. MOLL, and LOHL vs. HOLL), indicating that C18:2 is more potent than C18:1 for depressing milk fat. Increasing dietary FA content from no fat to LOLL, which increased primarily C18:1 and C18:2 with small increases in C18:0 and C16:0, decreased the secretion of C <16 but increased total C18 secretion in milk. This suggests that biohydrogenation intermediates act to decrease mammary FA synthesis at low levels of added C18:1 and C18:2. No significant monensin × fat interactions were detected for the milk composition parameters analyzed; however, a monensin × fat interaction was found for milk fat trans-10 C18:1 proportion.

Effects of monensin on metabolic parameters, feeding behavior, and productivity of transition dairy cows.

The effects of monensin on transition cow metabolism may be dependent on modulation of feeding behavior, rumen pH, and expression of key metabolic genes. Multiparous Holstein cows were used to determine the effects of monensin (400mg/cow daily) on these variables. Cows were randomly assigned, based on calving date, to control or monensin treatments (n = 16 per treatment) 21 d before their expected calving date, and cows remained on treatments through 21 d postpartum. Feeding behavior and water intake data were collected daily. Liver biopsies were conducted after assessing BCS and BW on d -21, -7, 1, 7, and 21 relative to calving for analysis of triglyceride (TG) content as well as mRNA abundance of cytosolic phosphoenolpyruvate carboxykinase, carnitine palmitoyltransferase 1a, and apolipoprotein B. Blood samples were collected 21, 7, and 4 d before expected calving and 1 (day of calving), 4, 7, 14, and 21 d postpartum for nonesterified fatty acid, ß-hydroxybutyrate, glucose, insulin, and haptoglobin analyses. Ruminal pH was collected every 5 min on d 1 through 6 postpartum via a wireless indwelling probe. On d 7 postpartum, a caffeine clearance test was performed to assess liver function. Data were analyzed using mixed models with repeated measures over time. Monensin decreased mean plasma ß-hydroxybutyrate (734 vs. 616 ± 41 µM) and peak concentrations (1,076 vs. 777 ± 70 µM on d 4 postpartum). Monensin also decreased time between meals prepartum (143 vs. 126 ± 5.0 min) and postpartum (88.8 vs. 81.4 ± 2.9 min), which was likely related to a smaller ruminal pH standard deviation in the first day after cows changed to a lactation ration (0.31 vs. 0.26 ± 0.015). Monensin also increased liver mRNA abundance of carnitine palmitoyltransferase 1a (0.10 vs. 0.15 ± 0.002 arbitrary units), which corresponded to a slower rate of liver TG accumulation from d -7 to +7 (412 vs. 128 ± 83 mg of TG/g of protein over this time period). No significant effects of monensin supplementation were observed on milk production, liver cytosolic phosphoenolpyruvate carboxykinase, apolipoprotein B, plasma nonesterified fatty acid, glucose, insulin, or haptoglobin. No effects on disease incidence were detected, but sample size was small for detecting such effects. Overall, results confirm that the effects of monensin on transition cows extend beyond altered propionate flux.

Feeding native rumen microbial supplements increases energy-corrected milk production and feed efficiency by Holstein cows.

The objective of this study was to investigate the effects of 2 novel native microbial feed supplements (MFS) on milk yield, milk composition, dry matter intake (DMI), body weight (BW), body condition score (BCS), and efficiency. A total of 73 lactating Holstein cows were enrolled over 4 mo. During a 2-wk covariate period, baseline data were collected. Using a randomized complete block design, cows were blocked in groups of 3 by days in milk (DIM), production, and parity. Within blocks cows were randomly assigned to a basal diet with a 150 g/d ground corn topdress and no MFS (control), the basal diet supplemented with a 5 g/d dose of Clostridium beijerinckii ASCUSDY20 and Pichia kudriavzevii ASCUSDY21 in a carrier mixed with a 150 g/d ground corn topdress (MFS1), or the basal diet with a 5 g/d dose of Ruminococcus bovis ASCUSDY10, Butyrivibrio fibrisolvens ASCUSDY19, Clostridium beijerinckii ASCUSDY20, and Pichia kudriavzevii ASCUSDY21 in a carrier mixed in a 150 g/d ground corn topdress (MFS2) for 140 d. Cows were fed the diets as a total mixed ration (TMR) for ad libitum DMI once daily. Analysis showed numerical increases in performance variables such as milk yield and fat and protein contents in both MFS1 and MFS2, although the differences were not significant over time. For energy-corrected milk (ECM), other solids percentage, and BW there were significant treatment by time interactions. Cows fed MFS1 and MFS2 produced 1.4 kg/d and 1.6 kg/d more ECM than control cows, respectively (time-dependent treatment significance). Up to wk 13 of treatment, both MFS groups had higher ECM production than the control group but after this point MFS1 rapidly decreased to the same level as control. All cows displayed a gradual decrease in ECM after wk 13 but the MFS2 group remained greater than both control and MFS1. A similar trend in the content of other milk solids was observed. Cow BW was affected over time; both control and MFS2 cows gained weight at a similar rate throughout the study, whereas the weight gain of MFS1 cows decreased briefly from wk 13 to 17 before increasing again. Feed efficiency tended to be improved by MFS, with greater improvement by MFS2 than by MFS1. The DIM when the supplementation began had a significant impact on MFS2 efficacy, where the supplementation of native rumen microbes at an earlier DIM resulted in a greater gain in ECM over the 140-d trial. A similar trend was observed for MFS1, although the effect was not significant. Our results suggest that dairy cow lactation stage and energy requirements also play an important role in product efficacy.

Postabsorptive carbohydrate adaptations to heat stress and monensin supplementation in lactating Holstein cows.

Multiparous cows (n=34, 89 d in milk, 537 kg) housed in environmental chambers were fed a control total mixed ration or one containing monensin (450 mg/cow per day) during 2 experimental periods (P): (1) thermal neutral (TN) conditions (constant 20°C) with ad libitum intake for 9 d, and (2) heat stress (HS, n=16) or pair-fed [PF; in TN (PFTN); n=18] for 9 d. Heat-stress was cyclical with temperatures ranging from 29.4 to 38.9°C. Rectal temperatures and respiration rates increased in HS compared with PFTN cows (38.4 to 40.4°C, 40 to 93 breaths/min). Heat stress reduced dry matter intake (DMI, 28%), and by design, PFTN cows had similar intakes. Monensin-fed cows consumed less DMI (1.59 kg/d) independent of environment. Milk yield decreased 29% (9.1 kg) in HS and 15% (4.5 kg) in PFTN cows, indicating that reduced DMI accounted for only 50% of the decreased milk yield during HS. Monensin had no effect on milk yield in either environment. Both HS and PFTN cows entered into calculated negative energy balance (-2.7 Mcal/d), and feeding monensin increased feed efficiency (7%) regardless of environment. The glucose response to an epinephrine (EPI) challenge increased (27%) during P2 for both HS and PFTN cows, whereas the nonesterified fatty acid response to the EPI challenge was larger (56%) during P2 in the PFTN compared with the HS cows. Compared with P1, whole-body glucose rate of appearance (Ra) decreased similarly during P2 in both HS and PFTN cows (646 vs. 514 mmol/h). Although having similar rates of glucose Ra, HS cows synthesized approximately 225 g less milk lactose; therefore, on a milk yield basis, glucose Ra decreased (3.3%) in PFTN but increased (5.6%) in HS cows. Regardless of environment, monensin-fed cows had increased (10%) glucose Ra per unit of DMI. From the results we suggest that the liver remains sensitive but adipose tissue becomes refractory to catabolic signals and that glucose Ra (presumably of hepatic origin) is preferentially utilized for processes other than milk synthesis during HS.

Effects of dietary microbial feed supplement on production efficacy in lactating dairy cows.

This study evaluated the effects of a microbial feed supplement (MFS; Galaxis, Ascus Biosciences Inc.) comprising 2 native rumen microbes on performance parameters in mid-lactation dairy cows. Forty-six lactating primiparous and multiparous Holstein cows [629 ± 62 kg of body weight, mean ± standard deviation (SD); parity 1.64 ± 0.49; 119 ± 38 days in milk; 45.11 ± 3.81 and 52.73 ± 4.77 kg/d of milk yield for primiparous and multiparous, respectively] were enrolled in a study containing 3 experimental periods (P). During all periods, enrolled cows were fed the same base total mixed ration (TMR) ad libitum once daily. During P1 (7 d), baseline data were obtained for covariate analysis. At the beginning of P2 (60 d), cows were assigned to 1 of 2 dietary treatment groups in a randomized complete block design to balance for milk yield (MY), parity, and days in milk: (1) a control diet (CON; base TMR; n = 23), or (2) a control diet supplemented with 5 g/d of MFS (MFS; n = 23). Sample size was determined based on previous, unpublished results involving this MFS; a 3-kg difference between groups with a SD of 3.5 kg could be detected with sufficient power (0.81) using a total sample size of 46 cows. Treatment was top-dressed and hand-mixed into the top one-third of the TMR. During P3 (7 d), no treatment was administered, and all cows were fed the base TMR. When analyzing all cows in the data set, MFS had little to no effect on performance. However, modeling revealed that the fixed effect of covariate milk production level had a significant effect on the response of MY and ECM, and further investigation of the data revealed that treatment effectiveness in P2 correlated with milk production during P1. Cows were retrospectively categorized into 2 milk production groups (MPG) balanced for parity: MPG1 (i.e., <53 kg/d of ECM during P1; n = 34) or MPG2 (i.e., ≥53 kg/d of ECM during P1; n = 12). Energy-corrected milk was increased by 4.4% in MFS-administered MPG1 cows compared with CON cows during P2. Although there were no significant effects of MFS on production variables for MPG2 cows, MY tended to be decreased by 3.9% in MFS-administered cows compared with CON cows. Further investigation is needed to understand production level response differences and the effect of supplemented native rumen microbes on animal health and productivity.

Lactation response of dairy cows receiving bovine somatotropin and fed rations varying in crude protein and undegradable intake protein.

Our experiment evaluated lactation and metabolic responses of Holstein cows injected with somidobove (recombinant bST) and fed one of four isocaloric rations containing either 14 (low) or 17% (high) CP and undegradable intake protein of 33 (low) or 40% (high) of CP. Multiparous cows (n = 37) in early lactation, averaging 37 kg/d of milk, received somidobove (640 mg per injection) at 28-d intervals for 112 d and one of four protein rations: low-low, low-high, high-low, and high-high. Nine other multiparous controls were fed low-low ration with no somidobove. On the low-low ration, somidobove significantly increased milk yield by 2.3 kg/d, but not 3.5% FCM (1.7 kg/d), intakes of DM or CP, or milk composition. Milk and 3.5% FCM increased by 1.7 and 2.1 kg/d in cows fed high undegradable intake protein but there was no effect on milk composition, BW, or DM intake. Ration CP had no effect on production variables in cows receiving somidobove. Serum urea was higher in cows fed high CP rations; undegradable intake protein was without effect. Plasma leucine was higher in cows fed high undegradable intake protein. Administration of somidobove to cows fed low-low rations reduced plasma methionine, serum albumin, hemoglobin, and albumin:globulin ratio. Milk production of high producing dairy cows receiving somidobove may be limited by the amount of protein available at the small intestine.

The effects of a sustained-release recombinant bovine somatotropin (somidobove) on udder health for a full lactation.

The lactational effect of somidobove (recombinant bST) injections on dairy cows in a full lactation was measured in 193 primiparous and 159 multiparous Holsteins. Experimental animals, distributed across six study sites, were administered a sustained-release formulation of somidobove by subcutaneous injection every 28 d beginning 36 to 49 DIM. Randomization at each site determined which of the following somidobove treatments cows received: 0 mg (control), 160 mg (primiparous only), 320 mg, 640 mg, or 960 mg (multiparous only). In addition to lactation response, cows on study were monitored for mastitis. Clinical mastitis was detected by examination of quarter foremilk at each milking. Milk from 300 of 352 cows was monitored for new IMI by routine collection and culture of duplicate quarter milk samples. Somatic cell counts were conducted on individual composite milk samples collected weekly throughout the experiment. No evidence existed of an association between somidobove administration and the incidence or duration of clinical mastitis. Furthermore, somidobove administration was not associated with an increase in new IMI or prevalence of infection by common mastitis pathogens or pathogen groups. Somatic cell counts were low in all treatment groups, but a dose-related trend was found for increased SCC in both primiparous and multiparous cows.

Vitamin D metabolites in plasma of cows fed a prepartum low-calcium diet for prevention of parturient hypocalcemia.

Our objective was to characterize changes in vitamin D metabolites of plasma in Jersey cows fed a prepartum low-calcium diet. Eight cows were fed a high-calcium diet (80 g/day) and eight were fed a low-calcium diet (8 g/day) at least 14 days before parturition. Calcium concentrations in plasma decreased after initiation of feeding either diet, but cows fed low-calcium diet tended to have lower prepartum calcium and phosphorus and greater peripartal calcium in plasma. Hydroxyproline in plasma was greater during peripartal period in cows fed low-calcium diet. Prepartum 1,25-dihydroxyvitamin D in plasma tended to be greater in cows fed low calcium. Increases in 1,25-dihydroxyvitamin D were only 2 and 3 days after initiation of the low-calcium diet; during the first 2 days after parturition, however, 1,25-dihydroxyvitamin D tended to be lower in those cows fed low calcium. As parturition neared, 24,25-dihydroxyvitamin D tended to be lower in cows fed the low calcium-diet. Usual early postpartum changes in calcium phosphorus, magnesium, 1,25-dihydroxyvitamin D, and hydroxyproline were seen during first few days after initiation of feeding low calcium. Thus, we propose that the preventative action of the low-calcium diet is associated with preparation of the calcium homeostatic mechanism several days before the calcium demand of initiation of lactation.

Lactation response of dairy cows receiving bovine somatotropin via daily injections or in a sustained-release vehicle.

Three experiments were conducted to characterize metabolic and milk production responses of dairy cows receiving recombinantly derived bovine somatotropin administered either by daily injection or in a sustained-release vehicle. In Experiment 1, somatotropin (25 mg/d) purified by two methods was given by daily injection for 14 d and resulted in 3.5 and 3.8 kg/d more milk than controls. Percentages of fat and total solids in milk were also increased by somatotropin. Eleven hematology indices and 12 metabolites, minerals, and enzyme activities in serum were unaffected by somatotropin. In Experiments 2 and 3, somatotropin was administered in a sustained-release vehicle during an 84-d treatment period. In Experiment 2, administration of 960 mg of somatotropin at 28-d intervals increased milk and SCM yields by 4.1 and 3.3 kg/d compared with yields of controls. There were no significant differences in other production parameters. In Experiment 3, 320, 640, and 960 mg somatotropin were each administered in the sustained-release vehicle at intervals of 14, 21, and 28 d. An uninjected group served as control. Cows receiving somatotropin averaged 3.5 to 5.9 kg/d more milk than controls across all injection intervals. Among doses, milk yield was greater at 960 mg than at 320 or 640 mg. There were no significant differences in milk or SCM among injection groups. These experiments demonstrate the comparable efficacy of somatotropin when given by daily injections or in a sustained-release vehicle.