Effect of withdrawing an immunomodulatory feed additive from lactating cow diets on peripheral blood mononuclear cell proliferation.

Supplementation of immunomodulatory feed additives, such as OmniGen AF (OG), helps to maintain immune competency; however, it is unknown if immune benefits persist in lactating cows after OG is removed from the diet. The aim of this trial was to evaluate the effect of withdrawing OG from the diet on peripheral blood mononuclear cells (PBMC) proliferation of midlactation dairy cows. Multiparous Holstein cows (N = 32), blocked by parity (2.7 ± 0.8) and days in milk (153 ± 39 d) were randomly assigned to one of the two dietary treatments within each block: diets were top dressed with either OG (56 g/d/cow) or placebo (CTL, 56 g/d/cow). Cows were housed in the same free-stall pen and individually fed once per day through Calan gates. All cows were fed the same diet containing OG for at least 1 yr before the onset of treatments. Cows were milked three time per day and milk yield was recorded at each milking. Milk samples were collected from three consecutive milkings weekly and composition analyzed. Body weight (BW) and condition score were measured weekly. Blood samples were collected at -1, 1, 3, 5, and 7 wk relative to the onset of treatments for the isolation of PBMC. The PBMC were cultured with concanavalin A (ConA) and lipopolysaccharides (LPS) for 72 h in vitro to determine proliferative responses. Prior to the experiment, cows in both treatments had similar disease incidence. During the experiment, cows did not show symptoms of disease. Withdrawing OG from the diet did not affect (P ≥ 0.20) milk yield or composition, intake, or BW. Compared with CTL, feeding OG maintained greater body condition score (2.83 vs. 2.92, P = 0.04). Regardless of time, relative to CTL, PBMC isolated from cows fed with OG had a greater proliferative rate when stimulated with LPS (stimulation index: 1.27 vs. 1.80, P = 0.05) and tended to have greater proliferation when stimulated with ConA (stimulation index: 5.24 vs. 7.80, P = 0.08). In conclusion, withdrawing OG from the diet of midlactation cows reduced proliferative response of PBMC suggesting that the immunomodulatory role of OG is lost as early as 1 wk after its withdrawal from the diet of lactating dairy cows.

We hypothesized that withdrawing an immune modulatory feed additive (OmniGen AF [OG]) from the diet would reduce immune function of lactating dairy cows. Thus, this short communication aimed to examine if withdrawing OG from the diet of the lactating dairy cow could affect her immune function. Thirty-two midlactation Holstein cows were fed OG prior to the experiment. In the 8 wk experiment, OG was removed from the diet of 16 cows, but remained in the diet of the other 16 cows. We observed that withdrawing OG did not affect the intake and milk production but reduced the ability of the circulating immune cells of the cow to proliferate when challenged with mitogens in vitro. These results suggest withdrawing OG from the diet reduces the immune function of lactating dairy cows, increasing the risk of developing diseases.

Effect of milk replacer feeding rate and frequency of preweaning dairy calves in the southeastern United States: Glucose metabolism.

The objective of this experiment was to examine the effect of milk replacer (MR) feeding rate (FR) and frequency (FF) on glucose metabolism before and after weaning during summer and winter in the subtropical climate of the southeastern United States. Holstein calves (n = 48/season) were enrolled at 8 d of age (DOA) in the summer (June to August, body weight = 40.6 ± 0.7 kg) and winter (November to January, body weight = 41.9 ± 0.8 kg). In each season, calves were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement including 2 FR [0.65 (low) or 0.76 kg of solids/d (high) of a 26% CP and 17% fat MR] and 2 FF [2× (0700 and 1600 h) or 3× (0700, 1600, and 2200 h)]. Calves were managed similarly and housed in polyethylene hutches bedded with sand. Milk replacer (12.5%) was fed based on treatments until 42 DOA when FR was reduced by half and offered 1×/d (0700 h) for 7 d. Plasma was collected weekly at 1400 h for analyses of glucose and insulin concentrations in all calves. Pre- and postprandial glucose and insulin concentrations of a subset of calves (n = 10/treatment per season) were measured on 20 DOA. A subset of calves (n = 8/treatment per season) was subjected to an intravenous glucose tolerance test (GTT) on 27 and 57 DOA and insulin challenge on 28 and 58 DOA at 1030 h. Average ambient temperature was 26.1 ± 2.2°C in summer and 12.9 ± 5.4°C in winter. During the preweaning period in both seasons, feeding high increased plasma glucose concentrations compared with low, and increasing FF reduced basal insulin concentrations. Compared with 2×, feeding 3× did not affect postprandial glucose but lowered insulin in the summer, whereas in the winter, increased glucose from 30 to 180 min but lowered insulin from 240 to 420 min after MR feeding. Following GTT before weaning in both seasons, 3× reduced insulin increment and area under the curve compared with 2× without affecting glucose disposal. After weaning, treatment did not affect glucose disposal or insulin responses after GTT during winter, but calves fed 3× had faster glucose disposal and stronger insulin responses than 2× during summer. In both summer and winter, preweaned calves fed 3× had greater decrement and area under the curve of plasma glucose after insulin challenge, suggesting enhanced peripheral tissue insulin response compared with 2×. This effect persisted after weaning only during summer. Increasing FR had no effect on metabolic responses in both seasons. In conclusion, increasing MR FF from 2 to 3 times per day reduced insulin secretion but enhanced insulin response on peripheral tissues of preweaned calves regardless of season.

Effects of milk replacer feeding rate and frequency of preweaning dairy calves in the southeastern United States: Performance, abomasal emptying, and nutrient digestibility.

To evaluate the effects of milk replacer (MR) feeding rate (FR) and frequency (FF) on performance, abomasal emptying, and nutrient digestibility in the southeastern United States, Holstein calves (n = 48/season) were enrolled at 8 d of age (DOA) during summer [June to August, body weight (BW; mean ± SD) = 40.71 ± 4.35 kg] and winter (November to January, BW = 42.03 ± 3.83 kg). Within season, calves were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement including 2 FR [0.65 (low) or 0.76 kg of solid per day (high) of a 26% crude protein and 17% fat MR], and 2 FF [2× (0700 and 1600 h) or 3× (0700, 1600, and 2200 h) daily]. Calves were housed in polyethylene hutches and managed similarly throughout the trial. Milk replacer (12.5% solids) was fed to calves based on their respective treatments until 42 DOA, when MR allowance was reduced by 50% and offered once a day (0700 h) for the following 7 d until weaning. Calves remained on trial until 63 DOA. Calf starter and water were offered ad libitum. Ambient temperature and relative humidity inside and outside hutches were measured hourly. Starter and MR intakes were recorded daily. Respiration rate and rectal temperature were recorded 3 times a week. Structural growth and BW were measured weekly. Acetaminophen (50 mg/kg of BW) mixed with MR was fed to a subset of calves (0700 h, n = 10/treatment per season) on 20 DOA. Plasma was collected at 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330, 360, 420, and 480 min after feeding, to analyze acetaminophen. The acetaminophen concentration-time curve was modeled to the first derivative of Siegel's modified power exponential equation, and the time for plasma acetaminophen to reach maximum (Tmax) was calculated to evaluate abomasal emptying rate. During the pre- (14.9-17.9 DOA) and postweaning (51.0-54.0 DOA) periods, a subset (n = 8/treatment per season) of calves was used to determine the apparent digestibility of nutrients, using chromic oxide as the external marker. Feeding 3× reduced preweaning respiration rate during summer and reduced rectal temperature during winter. Increasing FR improved BW gain and structural growth. Feeding more times per day tended to improve growth during winter but not summer. We found no effect of treatment on nutrient digestibility. Increasing FR had no effect on Tmax during winter but tended to delay Tmax of plasma acetaminophen during summer. Regardless of season, increasing FF lowered Tmax of plasma acetaminophen. In conclusion, increasing FF accelerated abomasal emptying and might reduce heat load of preweaning dairy calves but improved growth only during winter. Increased MR allowance improved growth in both seasons but delayed abomasal emptying only under heat stress conditions.

Impact of heat stress and a feed supplement on hormonal and inflammatory responses of dairy cows.

The aim of this trial was to evaluate the effects of an immunomodulatory supplement (OmniGen AF, OG; Phibro Animal Health Corp.) and heat stress on hormonal, inflammatory, and immunological responses of lactating dairy cows. Sixty multiparous Holstein cows were randomly assigned to 4 treatments in a 2 × 2 factorial arrangement using 2 environments: cooled using fans and misters, or noncooled, and 2 top-dressed feed supplements (56 g/d): OG or a placebo (CTL). Temperature-humidity index averaged 78 during the 8-wk trial. Blood was drawn to analyze cortisol, prolactin, and circulating tumor necrosis factor (TNF)-α and IL-10. Peripheral blood mononuclear cells (PBMC) were isolated and stimulated with hydrocortisone, prolactin, or lipopolysaccharide (LPS), individually or in several combinations, to assess induced proliferation and cytokine production. At d 52, 6 cows per treatment were injected i.v. with an LPS bolus (ivLPS) to assess hormone and cytokine responses. For cooled cows, feeding OG increased plasma cortisol concentration relative to CTL. Noncooled cows fed CTL had lower circulating TNF-α concentrations than noncooled-OG and cooled-CTL cows, with cooled-OG intermediate. Hydrocortisone+LPS-stimulated PBMC from OG cows tended to proliferate more than CTL. Relative to cooled cows, PBMC from noncooled cows produced more TNF-α and IL-10 when stimulated with LPS. Following ivLPS, cooled-OG cows had a greater cortisol response than the other treatments. In conclusion, OG supplementation enhanced cortisol release under basal condition and induced inflammation with cooling compared with CTL. This suggests that heat stress inhibits OG-mediated cortisol release. Heat stress seemed to enhance the inflammatory responses of PBMC from lactating cows. However, OG supplementation promoted PBMC proliferation under stress, or in the presence of hydrocortisone.

Response of lactating dairy cows fed different supplemental zinc sources with and without evaporative cooling to intramammary lipopolysaccharide infusion: metabolite and mineral profiles in blood and milk.

The objective of this study was to determine the effect of evaporative cooling and dietary supplemental Zn source on blood metabolites, insulin and mineral concentrations, and milk mineral concentrations following intramammary lipopolysaccharide (LPS) infusion. Seventy-two multiparous Holstein cows were assigned to one of four treatments with a 2 × 2 factorial arrangement. Treatments included two environments: with or without evaporative cooling using fans and misters over the freestall and feedbunk, and two dietary sources of supplemental Zn: 75 mg/kg of dry matter (DM) supplied by Zn hydroxychloride (inorganic Zn; IOZ) or Zn hydroxychloride (35 mg of Zn/kg of DM) + Zn-Met complex (ZMC; 40 mg of Zn/kg of DM). A subset of cows (n = 16; 263 ± 63 d in milk) was infused with 10 µg of LPS or a saline control in the left or right rear quarters on day 34 of the environmental treatment. Individual milk samples collected from LPS-infused quarters at -4, 0, 6, 12, 24, 48, 72, 96, and 144 h relative to infusion were analyzed for minerals. Blood samples were collected at the same time with an additional sample collected at 3 h post-infusion to analyze glucose, nonesterified fatty acids (NEFA), insulin, and minerals. Cooling by time interactions (P ≤ 0.07) were observed for plasma glucose, NEFA, and serum insulin. Compared with cooled cows, non-cooled cows had lower concentrations of plasma glucose except at 3 h following intramammary LPS infusion, greater serum insulin at 3 and 12 h, and lower plasma NEFA at 24 and 48 h after infusion. Relative to cooled cows, non-cooled cows tended (P = 0.07) to have lower serum K concentration and had lower (P < 0.01) serum Zn 6 h following infusion (cooling by time interaction: P < 0.01). Relative to ZMC cows, IOZ cows had greater (P ≤ 0.09) concentrations of plasma Se, skim milk Na and Se, and skim milk Na to K ratio. Regardless of treatment, intramammary LPS infusion reduced (P < 0.01) serum or plasma concentrations of Ca, Mg, Zn, Fe, and Se, but increased (P < 0.01) their concentration in skim milk. In conclusion, deprivation of cooling resulted in more rapid and prolonged insulin release and influenced the systemic and mammary mineral metabolism during mammary inflammation induced by LPS of lactating dairy cows. Dietary supplementation of Zn-Met complex reduced blood and milk Se concentrations compared with cows fed Zn from an inorganic source.

Short communication: Effect of supplemental zinc source with and without evaporative cooling on systemic and mammary metabolism of lactating dairy cows during summer.

The negative effects of heat stress partly result from disturbed systemic metabolic responses and possibly altered mammary gland metabolism of lactating dairy cows. Our previous research reported that supplemental dietary Zn sources may affect milk fat synthesis of lactating cows during summer. Thus, our objective was to evaluate the systemic and mammary metabolism of cows fed 2 supplemental Zn sources under 2 environmental conditions. Multiparous lactating Holstein cows (n = 72; days in milk: 99.7 ± 13.4 d; parity: 2.9 ± 0.3) were randomly assigned to 4 treatments in a 2 × 2 factorial arrangement. Treatments included 2 different environments: cooled (CL) using fans and misters or noncooled (NC), and 2 supplemental Zn sources: 75 mg of Zn hydroxychloride/kg of DM (IOZ) or 35 mg of Zn hydroxychloride/kg of DM + 40 mg of Zn-Met complex/kg of DM (ZMC). The 168-d experiment was divided into baseline and environmental challenge phases, 84 d each. During the baseline phase, all cows were cooled and fed respective dietary treatments, and during the environmental challenge phase cows continued receiving the same diets but NC cows were deprived of cooling. Temperature-humidity index averaged 77.6 ± 3.8 and 77.8 ± 3.8 for CL and NC pens, respectively, during the environmental challenge phase. Plasma was collected before the baseline phase and at 1, 3, 5, 12, 22, 26, 41, 54, 61, 68, 75, and 81 d of the environmental challenge phase for metabolites and insulin analyses. Mammary biopsies were collected before the baseline phase and at 7 and 56 d of the environmental challenge phase to measure mRNA abundance of proteins related to mammary metabolism. Compared with CL, NC reduced plasma glucose, nonesterified fatty acids, ß-hydroxybutyrate, and triglyceride concentrations, but increased insulin concentration. Cows fed ZMC had greater plasma triglyceride concentration than IOZ. Treatments had no effect on mRNA abundance of protein related to mammary fatty acid and glucose metabolism except that NC cows had greater mammary mRNA abundance of 6-phosphogluconate dehydrogenase and ATP-dependent 6-phosphofructokinase than CL cows. In conclusion, deprivation of evaporative cooling influenced the metabolism of lactating dairy cows but dietary Zn source had no apparent effect.

Impact of heat stress on lactational performance of dairy cows.

Lactating dairy cows exhibit a myriad of responses to heat stress. These responses partially facilitate the thermal balance between heat gain and heat loss, but also account for reduction in productivity. Decreased milk yield is the most recognized impact of heat stress on a dairy cow and results in significant economic loss to dairy producers. The reduced milk yield by heat stress is observed when daily average temperature-humidity index exceeds 68, above which the milk yield of a cow is negatively correlated with temperature-humidity index or dry bulb temperature. Milk yield is also positively correlated with body temperature of the cows under evaporative cooling, which reflects the positive relationship between metabolic heat production and milk yield. During summer, feed intake is positively correlated with milk yield, and the decreased intake explains at least half of the reduction in milk yield by heat stress. These emphasize the importance of maintaining intake on productivity during summer. Although not entirely clear, mechanisms that mediate the reduced milk yield by heat stress in addition to intake may be multifactorial. These could include but are not limited to altered metabolism, potential activation of immune system and inflammation, changes in behavior, and altered mammary gland development and function.

Response of lactating dairy cows fed different supplemental zinc sources with and without evaporative cooling to intramammary lipopolysaccharide infusion: intake, milk yield and composition, and hematologic profile1.

The objective of this study was to determine the effect of dietary supplemental Zn source and evaporative cooling on intake, milk yield and composition, and the rate of leukocyte migration into the mammary gland following intramammary lipopolysaccharide (LPS) infusion. Multiparous Holstein cows (n = 72) were assigned to one of four treatments with a 2×2 factorial arrangement including two sources of supplemental Zn: 75 mg/kg Zn hydroxychloride or 35 mg/kg Zn hydroxychloride + 40 mg/kg Zn-Met complex (ZMC) each with or without evaporative cooling. The cooling system was implemented by the use of fans and misters over the freestall and feeding areas. On day 34 of the experiment, cows (n = 16; days in milk = 263 ± 63 d) received an infusion of 10 µg of LPS, or a saline control, in the left or right rear quarters. Individual milk samples from both quarters were collected at -12, -4, 0, 6, 12, 24, 48, 72, 96, 120, 144, and 168 h relative to infusion and analyzed for composition and bovine serum albumin. Rectal temperature and respiration rate were assessed and blood samples were collected at the same time points (with an additional sample at 3 h) for analyses of lactose and cortisol. Complete blood counts were performed on samples collected within the first 24 h post infusion. Intramammary LPS infusion reduced (P < 0.01) milk yield, DMI and feed efficiency regardless of dietary or cooling treatments. Non-cooled cows tended (P = 0.09) to have greater feed efficiency (=milk yield/DMI) at 1 d after infusion than those subjected to cooling. Intramammary LPS infusion dramatically increased (P < 0.01) milk somatic cell count (SCC) but treatments had no apparent impact on milk SCC. Compared with cooled cows, non-cooled cows had greater (P < 0.05) plasma lactose concentrations, but lower (P < 0.03) blood concentrations of neutrophils and lymphocytes at 3 h post infusion. This suggests a greater leukocyte migration into the mammary gland of heat-stressed cows. In conclusion, noncooled cows tended to maintain greater feed efficiency and appeared to have greater leukocyte migration into the mammary gland immediately after intramammary LPS infusion compared with cooled cows. Dietary supplemental Zn source had no impact on measures assessed after intramammary LPS infusion.

PHYSIOLOGY SYMPOSIUM: Effects of heat stress during late gestation on the dam and its calf12.

Heat stress during late gestation in cattle negatively affects the performance of the dam and its calf. This brief exposure to an adverse environment before parturition affects the physiological responses, tissue development, metabolism, and immune function of the dam and her offspring, thereby limiting their productivity. During the dry period of a dairy cow, heat stress blunts mammary involution by attenuating mammary apoptosis and autophagic activity and reduces subsequent mammary cell proliferation, leading to impaired milk production in the next lactation. Dairy cows in early lactation that experience prepartum heat stress display reduced adipose tissue mobilization and lower degree of insulin resistance in peripheral tissues. Similar to mammary gland development, placental function is impaired by heat stress as evidenced by reduced secretion of placental hormones (e.g., estrone sulfate) in late gestation cows, which partly explains the reduced fetal growth rate and lighter birth weight of the calves. Compared with dairy calves born to dams that are exposed to evaporative cooling during summer, calves born to noncooled dry cows maintain lower BW until 1 yr of age, but display a stronger ability to absorb glucose during metabolic challenges postnatally. Immunity of the calves, both passive and cell-mediated immune function, is also impaired by prenatal heat stress, resulting in increased susceptibility of the calves to diseases in their postnatal life. In fact, dairy heifers born to heat-stressed dry cows without evaporative cooling have a greater chance leaving the herd before puberty compared with heifers born to dry cows provided with evaporative cooling (12.2% vs. 22.7%). Dairy heifers born to late-gestation heat-stressed dry cows have lower milk yield at maturity during their first and second lactations. Emerging evidence suggests that late-gestation heat stress alters the mammary gland microstructure of the heifers during the first lactation and exerts epigenetic alterations that might explain, in part, their impaired productivity.