Heat stress affects some physiological and productive variables and alters metabolism in dairy ewes

Heat stress (HS) has a significant economic impact on the global dairy industry. However, the mechanisms by which HS negatively affects metabolism and milk synthesis in dairy ewes are not well defined. This study evaluated the production and metabolic variables in dairy ewes under controlled HS conditions. Eight Lacaune ewes (75.5 ± 3.2 kg of body weight; 165 ± 4 d of lactation; 2.31 ± 0.04 kg of milk per day) were submitted to thermoneutral (TN) or HS conditions in a crossover design (2 periods, 21 d each, 6-d transition). Conditions (day-night, 12–12 h; relative humidity; temperature-humidity index, THI) were: TN (15–20°C; 50 ± 5%; THI = 59–65) and HS (28–35°C; 45 ± 5%; THI = 75–83). Ewes were fed ad libitum and milked twice daily. Rectal temperature, respiratory rate, feed intake, water consumption, and milk yield were recorded daily. Milk and blood samples were collected weekly. Additionally, TN and HS ewes were exposed to glucose tolerance test, insulin tolerance test, and epinephrine challenge. Heat stress reduced feed intake (?11%), and increased rectal temperature (+0.77°C), respiratory rate (+90 breaths/min), and water consumption (+28%). Despite the reduced feed intake, HS ewes produced similar milk to TN ewes, but their milk contained lower fat (?1.7 points) and protein (?0.86 points). Further, HS milk tended to contain more somatic cells (+0.23 log points). Blood creatinine was greater in HS compared with TN, but no differences in blood glucose, nonesterified fatty acids, or urea were detected. When glucose was infused, TN and HS had similar insulin response, but higher glucose response (+85%) was detected in HS ewes. Epinephrine infusion resulted in lower nonesterified fatty acids response (?215%) in HS than TN ewes. Overall, HS decreased feed intake, but milk production was not affected. Heat stress caused metabolic adaptations that included increased body muscle degradation and reduced adipose tissue mobilization. These adaptations allowed ewes to spare glucose and to avoid reductions in milk yield.     

The effects of heat stress on protein metabolism in lactating Holstein cows

Heat stress (HS) decreases milk protein synthesis beyond what would be expected based on the concomitant reduction in feed intake. The aim of the present study was to evaluate the direct effects of HS on milk protein production. Four multiparous, lactating Holstein cows (101 ± 10 d in milk, 574 ± 36 kg of body weight, 38 ± 2 kg of milk/d) were individually housed in environmental chambers and randomly allocated to 1 of 2 groups in a crossover design. The study was divided into 2 periods with 2 identical experimental phases (control phase and trial phase) within each period. During phase 1 or control phase (9 d), all cows were housed in thermal neutral conditions (TN; 20°C, 55% humidity) and fed ad libitum. During phase 2 or treatment phase (9 d), group 1 was exposed to cyclical HS conditions (32 to 36°C, 40% humidity) and fed ad libitum, whereas group 2 remained in TN conditions but was pair-fed (PFTN) to their HS counterparts to eliminate the confounding effects of dissimilar feed intake. After a 30-d washout period in TN conditions, the study was repeated (period 2), inverting the environmental treatments of the groups relative to period 1: group 2 was exposed to HS and group 1 to PFTN conditions. Compared with PFTN conditions, HS decreased milk yield (17.0%), milk protein (4.1%), milk protein yield (19%), 4% fat-corrected milk (23%), and fat yield (19%). Apparent digestibility of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, crude protein, and ether extract was increased (11.1–42.9%) in HS cows, as well as rumen liquor ammonia (before feeding 33.2%; after feeding 29.5%) and volatile fatty acid concentration (45.3%) before feeding. In addition, ruminal pH was reduced (9.5 and 6% before and after feeding, respectively) during HS. Heat stress decreased plasma free amino acids (AA; 17.1%) and tended to increase and increased blood, urine, and milk urea nitrogen (17.2, 243, and 24.5%, respectively). Further, HS cows had reduced plasma glucose (8%) and nonesterified fatty acid (39.8%) concentrations compared with PFTN controls. These data suggest that HS increases systemic AA utilization (e.g., decreased plasma AA and increased nitrogen excretion), a scenario that limits the AA supply to the mammary gland for milk protein synthesis. Furthermore, the increase in AA requirements during HS might represent the increased need for gluconeogenic precursors, as HS is thought to prioritize glucose utilization as a fuel at the expense of nonesterified fatty acids.     

Increased dietary vitamin D3 and calcium partially alleviate heat stress symptoms and inflammation in lactating Holstein cows independent of dietary concentrations of vitamin E and selenium

Twelve multiparous Holstein cows (42.2 ± 5.6 kg of milk/d; 83 ± 27 d in milk) were used in a split-plot design testing the effects of mineral and vitamin supplementation on the time course of animal performance, metabolism, and inflammation markers during heat stress. The main plot was the average concentrations of dietary vitamin E and Se (adequate: 11.1 IU/kg of vitamin E and 0.55 mg/kg of Se, and high: 223 IU/kg of vitamin E and 1.8 mg/kg of Se, respectively). Within each plot, cows were randomly assigned to (1) heat stress (HS) with adequate concentrations of vitamin D₃ and Ca (1,012 IU/kg and 0.73%, respectively), (2) HS with high concentrations of vitamin D₃ and Ca (HS+D₃/Ca; 3,764 IU/kg and 0.97%, respectively), or (3) pair-feeding (PF) in thermoneutrality with adequate concentrations of vitamin D₃ and Ca (1,012 IU/kg and 0.73% Ca) in a Latin square design with 14-d periods and 7-d washouts. The highest rectal temperature was recorded at 1700 h for HS (39.4°C; mean of d 1 to 14), being 1.2 and 0.8°C greater than for PF and HS+D₃/Ca, respectively. Respiratory rate and water intake were higher in HS (73 breaths/min and 115 L/d, respectively) relative to PF (28 breaths/min and 76 L/d). Heat stress decreased dry matter intake progressively, reaching a nadir on d 5 to 7 (33% reduction) and was not different between treatments. Milk yield decreased progressively in all treatments, but remained greater in PF relative to HS from d 3 to 14 (10%), whereas HS and HS+D₃/Ca were not different. Milk fat, protein, and lactose concentrations and yields were lower in HS relative to PF from d 3 to 14, but not different between HS and HS+D₃/Ca. Relative to PF, preprandial insulin concentrations were increased in HS, whereas plasma nonesterified fatty acids were decreased on d 7 and 14. Plasma lipopolysaccharide-binding protein concentrations increased in HS cows on d 7 and 14, respectively, relative to PF, whereas they were reduced in HS + D₃/Ca on d 14. Plasma C-reactive protein, tumor necrosis factor-α, and fecal calprotectin were increased in HS relative to both PF and HS+D₃/Ca on d 7 and 14. Rectal temperature was positively associated with plasma lipopolysaccharide-binding protein (r = 0.72), tumor necrosis factor-α (r = 0.74), C-reactive protein (r = 0.87), and with milk somatic cells (r = 0.75). Plasma 8-hydroxy-2-deoxyguanosine concentrations presented a 3-way interaction, where 8-hydroxy-2-deoxyguanosine was lower in HS than in PF on d 7 and 14, and lower in HS+D₃/Ca relative to HS on d 14 in the adequate vitamin E and Se treatment, but no effects were observed in the high vitamin E and Se group. Plasma superoxide dismutase concentrations increased over time, and were higher in HS relative to PF on d 14, whereas HS+D₃/Ca was similar to HS. Heat stress markedly reduced milk production and milk components while increasing markers of leaky gut and inflammation. In contrast, vitamin D₃ and Ca supplementation reduced hyperthermia (d 7–14), markers of leaky gut, and inflammation independent of dietary concentrations of vitamin E and Se.     

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.     

Rumen-protected methionine during heat stress alters mTOR,insulin signaling,and 1-carbon metabolism protein abundance in liver,and whole-blood transsulfuration pathway genes in Holstein cows

We investigated effects of rumen-protected Met (RPM) during a heat stress (HS) challenge on (1) hepatic abundance of mTOR, insulin, and antioxidant signaling proteins, (2) enzymes in 1-carbon metabolism, and (3) innate immunity. Holstein cows (n = 32; mean ± standard deviation, 184 ± 59 d in milk) were randomly assigned to 1 of 2 environmental groups, and 1 of 2 diets [total mixed ration (TMR) with RPM (Smartamine M; 0.105% dry matter as top-dress) or TMR without (CON); n = 16/diet] in a split-plot crossover design. There were 2 periods with 2 phases. During phase 1 (9 d), all cows were in thermoneutral conditions (TN; temperature-humidity index = 60 ± 3) and fed ad libitum. During phase 2 (9 d), half the cows (n = 8/diet) were exposed to HS using electric heat blankets. The other half (n = 8/diet) remained in TN, but was pair-fed to HS counterparts. After a 14-d washout and 7-d adaptation period, the study was repeated (period 2) and environmental treatments were inverted relative to phase 2, but dietary treatments were the same. Blood was collected on d 6 of each phase 2 to measure immune function and isolate whole-blood RNA. Liver biopsies were performed at the end of each period for cystathione β-synthase (CBS) and methionine adenosyltransferase activity, glutathione concentration, and protein abundance. Data were analyzed using PROC MIXED in SAS. Abundance of CUL3, inhibitor of antioxidant responses, tended to be downregulated by HS suggesting increased oxidative stress. Heat-shock protein 70 abundance was upregulated by HS. Phosphorylated mTOR abundance was greater overall with RPM, suggesting an increase in pathway activity. An environment × diet (E × D) effect was observed for protein kinase B (AKT), whereas there was a tendency for an interaction for phosphorylated AKT. Abundance of AKT was upregulated in CON cows during HS versus TN, this was not observed in RPM cows. For phosphorylated AKT, tissue from HS cows fed CON had greater abundance compared with all other treatments. The same effect was observed for EIF2A (translation initiation) and SLC2A4 (insulin-induced glucose uptake). An E × D effect was observed for INSR due to upregulation in CON cows during HS versus TN cows fed CON or RPM. There was an E × D effect for CBS, with lower activity in RPM versus CON cows during HS. The CON cows tended to have greater CBS during HS versus TN. An E × D effect was observed for methionine adenosyltransferase, with lower activity in RPM versus CON during HS. Although activity increased in CON during HS versus TN, RPM cows tended to have greater activity during TN. Neutrophil and monocyte oxidative burst and monocyte phagocytosis decreased with HS. An (E × D) effect was observed for whole-blood mRNA abundance of CBS, SOD1 and CSAD; RPM led to upregulation during TN versus HS. Regardless of diet, CDO1, CTH, and SOD1 decreased with HS. Although HS increased hepatic HSP70 and seemed to alter antioxidant signaling, feeding RPM may help cows maintain homeostasis in mTOR, insulin signaling, and 1-carbon metabolism. Feeding RPM also may help maintain whole-blood antioxidant response during HS, which is an important aspect of innate immune function.     

Effects of source on bioavailability of selenium,antioxidant status,and performance in lactating dairy cows during oxidative stress-inducing conditions

In the current study, we used heat stress (HS) as an oxidative stress model to examine the effects of hydroxy-selenomethionine (HMSeBA), an organic selenium source, on selenium's bioavailability, antioxidant status, and performance when fed to dairy cows. Eight mid-lactation Holstein dairy cows (141 ± 27 d in milk, 35.3 ± 2.8 kg of milk/d, parity 2 or 3) were individually housed in environmental chambers and randomly assigned to 1 of 2 treatments: inorganic Se supplementation (sodium selenite; SS; 0.3 mg of Se/kg of dry matter; n = 4) or HMSeBA supplementation (0.3 mg of Se/kg of dry matter; n = 4). The trial was divided into 3 continuous periods: a covariate period (9 d), a thermal neutral (TN) period (28 d), and a HS period (9 d). During the covariate and TN periods, all cows were housed in TN conditions (20°C, 55% humidity). During HS, all cows were exposed to cyclical HS conditions (32–36°C, 40% humidity). All cows were fed SS during the covariate period, and dietary treatments were implemented during the TN and HS periods. During HS, cows fed HMSeBA had increased Se concentrations in serum and milk, and total Se milk-to-serum concentration ratio compared with SS controls. Superoxide dismutase activity did not differ between Se sources, but we noted a treatment by day interaction in glutathione peroxidase activity as HS progressively reduced it in SS controls, whereas it was maintained in HMSeBA cows. Supplementation with HMSeBA increased total antioxidant capacity and decreased malondialdehyde, hydrogen peroxide, and nitric oxide serum concentrations compared with SS-fed controls. We found no treatment effects on rectal temperature, respiratory rate, or dry matter intake. Supplementing HMSeBA tended to increase milk yield and decrease milk fat percentage. No other milk composition parameters differed between treatments. We observed no treatment effects detected on blood biochemistry, except for a lower alanine aminotransferase activity in HMSeBA-fed cows. These results demonstrate that HMSeBA supplementation decreases some parameters of HS-induced oxidative stress.     

Rumen lipopolysaccharide and inflammation during grain adaptation and subacute ruminal acidosis in steers

Three rumen-fistulated Jersey steers were gradually adapted to a wheat-barley concentrate over a 4-wk period. Adaptation steps consisted of four 1-wk periods during which steers were fed diets with forage-to-concentrate (F:C) ratios of 100:0, 79:21, 59:41, and 39:61. The forage consisted of chopped hay (CH), and the concentrate consisted of pelleted concentrate containing 50% ground wheat and 50% ground barley. Steers were fed the all-forage diet ad libitum during wk 1. Feed offered in wk 2 to 4 was kept constant at the ad libitum intake during wk 1. On 2 d that were set 3 d apart during wk 5, subacute ruminal acidosis (SARA) was induced in the steers by feeding a diet with an F:C ratio of 24:76 by offering them 0.9 kg of CH at 0900 h followed by 2 meals of 3.0 kg each of wheat-barley pellets (WBP) at 1100 h and 1300 h and 0.9 kg of CH at 1700 h, to depress rumen pH for at least 3 h/d below 5.6. The average concentrate inclusion for the SARA induction diet was 76 ± 10% DM. During stepwise adaptation, time with pH below 5.6 increased to an average of 121 min/d when the steers were consuming a diet containing 61% DM as WBP. Dietary inclusion of WBP at the rate of 76% DM induced SARA because the steers spent an average of 219 min/d with pH below 5.6. The free ruminal lipopolysaccharide (LPS) concentration increased from 6,310 endotoxin units (EU)/mL with the all-forage diet to 18,197 EU/mL with the 61% concentrate diet. The ruminal LPS concentration increased to 26,915 EU/mL when SARA was induced. Serum haptoglobin increased from 0.53 mg/mL when steers were on the all-forage diet to 1.90 mg/mL with the 61% concentrate diet and were not increased further by inducing SARA. The serum amyloid-A concentration was not affected by increasing dietary concentrate during stepwise adaptation to the concentrate, but increased from 71 to 163 μg/mL when SARA was induced. A gradual increase in dietary concentrate so that the F:C ratio decreased to 39:61 resulted in increased ruminal LPS concentrations. Subsequent induction of SARA further increased ruminal LPS and activated an inflammatory response.     

Effects of dietary zinc source on the metabolic and immunological response to lipopolysaccharide in lactating Holstein dairy cows

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.     

Effects of dietary organic acid and pure botanical supplementation on growth performance and circulating measures of metabolic health in Holstein calves challenged by heat stress

To evaluate the effects of heat stress environmental conditioning and dietary supplementation with organic acid and pure botanicals (OA/PB) on growth in dairy calves, we enrolled 62 bull (noncastrated) and heifer calves in a study with a completely randomized design. Calves were assigned to 1 of 5 groups (n = 11 to 14/group): (1) thermoneutral conditions (TN-Con), (2) HS conditions (HS-Con), (3) thermoneutral conditions and pair-fed to match nutrient intake with HS-Con (TN-PF), (4) HS with low-dose OA/PB [75 mg/kg of body weight (BW); 25% citric acid, 16.7% sorbic acid, 1.7% thymol, 1.0% vanillin, and 55.6% triglyceride; HS-Low], or (5) HS with high-dose OA/PB (150 mg/kg of BW; HS-High). Supplements were delivered as a twice-daily bolus via the esophagus from wk 1 through 13 of life; all calves, including those on the control treatments, received an equivalent amount of triglyceride used for microencapsulation. Calves were raised in TN conditions from birth until weaning. After weaning, calves (62 ± 2 d; 91 ± 10.9 kg of BW) were transported to a new facility and remained in TN conditions [temperature-humidity index (THI): 60 to 69] for a 7-d covariate period. Thereafter, calves remained in TN or were moved to HS conditions (THI: diurnal change 75 to 83 during night and day, respectively) for 19 d. Clinical assessments were performed thrice daily, BW was recorded weekly, and blood was sampled on d 1, 2, 3, 8, 15, and 19. Upon experiment completion, calves from HS-Con and TN-Con were euthanized, and hot carcass and visceral organ weights were recorded. The mixed model included calf as a random effect; treatment, day, hour (when appropriate) as fixed effects, and the interactions of treatment × day and treatment × hour (when appropriate). Rectal and skin temperatures and respiration rates were greater in HS-Con than in TN-Con. During heat stress exposure, dry matter intake (DMI), average daily gain (ADG), and gain to feed (G:F) were lower in HS-Con relative to TN-Con. Comparing HS-Con and TN-PF, ADG and G:F were similar. Plasma fatty acid concentrations were elevated in TN-PF compared with HS-Con and TN-Con. Despite tendencies for increased aspartate aminotransferase, HS conditions did not overtly influence liver and inflammation markers. Liver weights were lower in HS-Con relative to TN-Con. During the first week of heat exposure, DMI was greater for HS-Low relative to HS-Con. Supplementation of OA/PB at low and high levels had a similar G:F to HS-Con. We conclude that reductions in DMI accounted for production losses during HS conditioning and that dietary OA/PB supplementation was not able to improve growth performance in heat-stressed calves.     

Effects of late-gestation heat stress independent of reduced feed intake on colostrum,metabolism at calving,and milk yield in early lactation of dairy cows

The objective of this study was to differentiate the effects of acute heat stress (HS) from those of decreased dry matter intake (DMI) during the prepartum period on metabolism, colostrum, and subsequent production of dairy cows. Holstein dairy cows (n = 30) with similar parity and body weight were randomly assigned to 1 of 3 treatments on 45 d before calving: (1) cooled (CL, n = 10) conditions with ad libitum feed intake, (2) HS conditions with ad libitum feed intake (n = 10), and (3) pair-fed cooled (CLPF, n = 10) with reduced DMI similar to the HS group while housed under cooled conditions. The reduction in the amount of feed offered to the CLPF cows was calculated daily as the percentage decrease from the average DMI of HS cows relative to the CL cows. For CLPF and CL cows, barns provided shade, sprinklers, and fans, whereas the HS cows were provided only with shade. Cows in all groups received individually the same total mixed ration. Cows were dried off 60 d before the expected calving. Cows in the HS group and, by design, the CLPF cows had reduced DMI (~20%) during the experiment. Heat stress decreased gestation length, first colostrum yield, and calf birth weight compared with CL and CLPF cows. Milk yield decreased 21% (5 kg) in the HS and 8% (2 kg) in CLPF cows, indicating that reduced feed intake during late gestation accounted for 60% of the total reduced milk yield. The CLPF cows exhibited an elevated NEFA concentration compared with the CL and HS cows. The HS cows had a greater mRNA abundance of HSP70 in the peripheral blood leukocytes at 21 d prepartum compared with the other groups. At calving, the mRNA abundance of HSP70 was greater in HS cows, followed by CLPF, compared with the CL cows. In conclusion, HS during the late gestation period caused metabolism and production differences, which were only partially attributed to reduced feed intake in dairy cows.     

Technical note: Developing a heat stress model in dairy cows using an electric heat blanket

Precisely studying the biological consequences of heat stress (HS) in agriculturally relevant animals typically requires expensive climate-controlled facilities, infrastructure inaccessible to most researchers. Thus, study objectives were to explore the efficacy of an electric heat blanket (EHB) as an alternative method for evaluating HS and to determine whether EHB-induced hyperthermia affects production parameters similar to natural HS. Lactating Holstein cows (n = 8; 133 ± 3 d in milk; 709 ± 31 kg; 2.6 ± 0.3 parity) were housed in individual box stalls and allowed to acclimate for 3 d. After acclimation, the trial consisted of 2 experimental periods (P). During P1 (3 d), cows were housed in thermoneutral conditions for collecting baseline data. During P2 (7 d), cows were fitted with an EHB. During the entire experiment cows were fed ad libitum, and dry matter intake (DMI) was recorded daily. Cows were milked twice daily (0600 and 1800 h), and milk samples were collected on d 2 and 3 of P1 and d 3 and 7 of P2. Rectal temperature, respiration rate, heart rate, and skin temperature were obtained twice daily (0600 and 1800 h) during both P1 and P2. Overall, there was an increase in rectal temperature and respiration rate at 0600 h (1.0°C and 25 breaths/min, respectively) and 1800 h (1.2°C and 29 breaths/min, respectively) during P2. The EHB decreased DMI and milk yield (25 and 21%, respectively) by the end of P2. During P2, milk protein tended to decrease (4.4%) compared with P1. In contrast, milk urea nitrogen increased (33%) during P2 relative to P1. No other differences were observed in milk composition. In summary, our results indicate that employing an EHB affects physiological and production parameters similarly to natural HS (i.e., increased rectal temperature and respiration rate, decreased DMI and milk yield); thus, the EHB is an effective and inexpensive research tool for evaluating the biological consequences of HS in lactating dairy cows.     

Feed additives containing sequestrant clay minerals and inactivated yeast reduce aflatoxin excretion in milk of dairy cows

The objective was to evaluate the efficacy of 2 dietary mycotoxin sequestrants, Toxy-Nil (TN) or Unike Plus (UP), in reducing aflatoxin (AF) M₁ concentrations in milk of dairy cows challenged with dietary AF. Thirty-two mid-lactation Holstein cows were blocked by parity, days in milk, and milk yield and were randomly assigned within block to receive one of the following treatments: (1) 2.8 mg of AF/cow per d (positive control, PC), (2) 2.8 mg of AF + 100 g of TN/cow per d, (3) 2.8 mg of AF + 100 g of UP/cow per d, or (4) no AF and no additives (negative control, NC). For 7 d, treatments, dispersed in 150 g of sweet feed carrier, were top-dressed twice daily by mixing into the top portion of the TMR at each feeding. After the experimental period, cows were fed the NC diet and clearance of AFM₁ via milk was monitored for 7 d. Feed and water were available ad libitum throughout the trial. Treatments had no effect on feed intake, milk yield, milk composition, or milk somatic cell count. Relative intake of AF was similar among PC, TN, and UP, averaging 106.5, 107.6, and 102.5 ± 2.9 μg/kg of diet dry matter, respectively. Relative intake of mycotoxin sequestrants was similar between TN and UP, averaging 0.4 and 0.4 ± 0.1% of diet dry matter, respectively. Concentration and mass of AFM₁ secreted in milk and in urine were similar between TN and UP, but were lower than PC; concentrations in milk averaged 0.2, 0.3, and 0.6 ± 0.1 μg/kg, respectively, and mass secreted in milk averaged 8.1, 9.8, and 20.5 ± 1.7 μg/d. Concentrations in urine averaged 6.9, 7.4, and 14.2 ± 1.5 μg/L, respectively, and mass secreted in urine averaged 225.7, 250.8, and 521.6 ± 53.1 μg/d. Likewise, concentration and mass of free AF excreted in feces were similar between TN and UP, but were lower than PC; concentrations averaged 7.7, 8.9, and 12.4 ± 0.6 μg/kg, respectively, and mass excreted averaged 57.8, 69.6, and 95.6 ± 4.8 μg/d. Transfer of AF from feed to AFM₁ in milk was reduced by 63 and 52%, and in urine, by 57 and 52% for TN and UP, respectively. Transfer of AF from feed to free AF in feces was reduced by 38 and 26% for TN and UP, respectively. The clearance rate of AFM₁ in milk did not differ among PC, TN, and UP (46.1, 66.5, and 50.0 ± 6.7%/d, respectively). Results indicate that dietary inclusion of 100 g of TN or UP significantly reduced AFM₁ in milk of cows consuming TMR containing approximately 105 μg of AF/kg of diet dry matter. Results also suggest that both TN and UP reduced absorption of AF.     

Effects of feed intake and thermal stress on mammary blood flow and other physiological measurements in lactating dairy cows

Six midlactation Holstein cows were exposed to treatments of thermal comfort environments with ad libitum or restricted (70% of ad libitum) DM intake and a thermal stress environment with ad libitum intake in two balanced 3 X 3 Latin squares to evaluate effects on mammary blood flow and other physiological measurements. Daily DM intake decreased from 17.8 kg in thermal comfort with ad libitum intake to 12.5 kg in thermal comfort with restricted intake and to 14.8 kg in thermal stress. Daily milk production decreased from 23.9 kg in thermal comfort ad libitum to 22.2 kg in thermal comfort restricted and 21.6 kg in thermal stress. Mammary blood flows (half udder) over the entire sampling interval (18 h) were 5.1, 4.3, and 4.5 L/min for treatments: thermal comfort, ad libitum intake; thermal comfort, restricted intake; and thermal stress, ad libitum intake. Mammary blood flows did not differ significantly among between treatments. Relationship of hald udder blood flow (L/min) to previous day's DM intake (kg/d) was described by the linear equation: L/min = .49 + (.27 kg/d); r2 = .46; and the quadratic equation: L/min = 6.04 - (.54 kg/d) + (.03 [kg/d]2); r2 = .55. Mammary blood flows 10 min before through 10 min after milking machine attachment were greater for cows at thermal comfort and ad libitum intake than for cows at thermal comfort and restricted intake or thermal stress and ad libitum intake (5.5, 4.8, and 4.8 L/min).     

Effects of feeding propionate and calcium salts of long-chain fatty acids on transition dairy cow performance

Multiparous Holstein cows (n = 40) were used in a randomized complete block design to determine the effects of feeding Ca and Na salts (1:1, wt/wt) of propionate and Ca salts of long-chain fatty acids (LCFA) on transition cow performance. All cows were fed the same basal diet once daily for ad libitum intake. Treatments (g/d) were 320 cornstarch (CS) as a control, 120 propionate (PRO), 120 propionate and 93 LCFA (PF1), and 178 propionate and 154 LCFA (PF2). Treatments were hand-mixed into the upper one-third of the TMR from 2 wk pre- through 3 wk postpartum. Intakes were recorded from 21 d pre- through 21 d postpartum. Energy density and crude protein were 1.54 and 1.65 Mcal/kg and 14.4 and 18.8% for pre- and postpartum diets, respectively. All cows received a common diet from 22 to 70 days in milk (DIM). Milk composition was analyzed on d 7, 14, and 21. Blood was sampled at 14, 7, and 2 d prepartum and 2, 7, 14, and 21 DIM. Pre- and postpartal dry matter intake (DMI) averaged 11.9 and 16.4 kg/d, respectively, and did not differ among treatments. A diet x week interaction for postpartal DMI was observed as cows fed PF2 consumed 2 kg/d less DM during wk 2 relative to other treatments. Milk yields from 22 to 70 DIM were 48.8, 48.5, 47.8, and 51.3 kg/d for CS, PRO, PF1, and PF2, respectively, and were not significantly affected by treatments. Milk true protein (3.32 vs. 3.16%) was increased and MUN (12.5 vs. 14.4 mg/dL) was decreased for CS relative to other treatments. Milk fat yield from cows fed PRO tended to be greater than those fed PF1 (1.58 vs. 1.29 kg/d). Plasma glucose, insulin, and beta-hydroxybutyrate were not affected by treatments. The PF2 treatment tended to decrease NEFA in plasma relative to PF1 over all times measured (492 and 670 muEq/L) and significantly decreased plasma NEFA relative to those fed PF1 postpartum (623 and 875 muEq/L). Relative to PF1, feeding propionate and LCFA at the higher level in this experiment improved energy balance postpartum as evidenced by decreased concentrations of plasma NEFA.     

Short communication: Antimethanogenic effects of 3-nitrooxypropanol depend on supplementation dose,dietary fiber content,and cattle type

3-Nitrooxypropanol (NOP) is a promising methane (CH₄) inhibitor. Recent studies have shown major reductions in CH₄ emissions from beef and dairy cattle when using NOP but with large variation in response. The objective of this study was to quantitatively evaluate the factors that explain heterogeneity in response to NOP using meta-analytical approaches. Data from 11 experiments and 38 treatment means were used. Factors considered were cattle type (dairy or beef), measurement technique (GreenFeed technique, C-Lock Inc., Rapid City, SD; sulfur hexafluoride tracer technique; and respiration chamber technique), dry matter (DM) intake, body weight, NOP dose, roughage proportion, dietary crude protein content, and dietary neutral detergent fiber (NDF) content. The mean difference (MD) in CH₄ production (g/d) and CH₄ yield (g/kg of DM intake) was calculated by subtracting the mean of CH₄ emission for the control group from that of the NOP-supplemented group. Forest plots of standardized MD indicated variable effect sizes of NOP across studies. Compared with beef cattle, dairy cattle had a much larger feed intake (22.3 ± 4.13 vs. 7.3 ± 0.97 kg of DM/d; mean ± standard deviation) and CH₄ production (351 ± 94.1 vs. 124 ± 44.8 g/d). Therefore, in further analyses across dairy and beef cattle studies, MD was expressed as a proportion (%) of observed control mean. The final mixed-effect model for relative MD in CH₄ production included cattle type, NOP dose, and NDF content. When adjusted for NOP dose and NDF content, the CH₄-mitigating effect of NOP was less in beef cattle (?22.2 ± 3.33%) than in dairy cattle (?39.0 ± 5.40%). An increase of 10 mg/kg of DM in NOP dose from its mean (123 mg/kg of DM) enhanced the NOP effect on CH₄ production decline by 2.56 ± 0.550%. However, a greater dietary NDF content impaired the NOP effect on CH₄ production by 1.64 ± 0.330% per 10 g/kg DM increase in NDF content from its mean (331 g of NDF/kg of DM). The factors included in the final mixed-effect model for CH₄ yield were ?17.1 ± 4.23% (beef cattle) and ?38.8 ± 5.49% (dairy cattle), ?2.48 ± 0.734% per 10 mg/kg DM increase in NOP dose from its mean, and 1.52 ± 0.406% per 10 g/kg DM increase in NDF content from its mean. In conclusion, the present meta-analysis indicates that a greater NOP dose enhances the NOP effect on CH₄ emission, whereas an increased dietary fiber content decreases its effect. 3-Nitrooxypropanol has stronger antimethanogenic effects in dairy cattle than in beef cattle.     

Effects of dietary antioxidant supplementation on metabolism and inflammatory biomarkers in heat-stressed dairy cows

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.     

Alterations in expression of gluconeogenic genes during heat stress and exogenous bovine somatotropin administration

Study objectives were to evaluate hepatic gluconeogenic enzyme gene expression in recombinant bovine somatotropin (rbST)-treated lactating dairy cattle during heat stress (HS) or in thermal-neutral, pair-fed (PF) animals. Twenty-two multiparous (99 d in milk, 656 kg of BW) Holstein cows were subjected to 3 consecutive experimental periods (7 d each): (1) thermal neutral, (2) HS or PF, and (3) HS or PF with rbST (Posilac, administered on d 1 of period 3). Liver biopsies were obtained on the final day of each period. Heat stress conditions progressively decreased dry matter intake for the first 5 to 6 d during period 2 before stabilizing (a decrease of 6.15 kg; 30%) on d 6 and 7, and feed intake remained stable and not different from period 2 during period 3. Cytosolic phosphoenolpyruvate carboxykinase mRNA abundance increased during PF, but was unaffected by HS or bST. Pyruvate carboxylase gene expression increased during HS and PF, and administrating bST decreased pyruvate carboxylase mRNA abundance during both HS and PF. Insulin-like growth factor-I gene expression increased following bST administration during HS and PF, confirming hepatic bST responsiveness. Exposure to HS leads to a change in hepatic gluconeogenic enzyme profile that appears to be dependent on plane of nutrition.     

An evaluation of an immunomodulatory feed ingredient in heat-stressed lactating Holstein cows: Effects on hormonal,physiological, and production responses

Holstein cows (n = 30) were balanced by days in milk, milk production, and parity (91 ± 5.9 d in milk, 36.2 ± 2.5 kg/d, and 3.1 ± 1.4, respectively) and fed OmniGen-AF (OG; Phibro Animal Health, Teaneck, NJ), an immune stimulant, at 0 g/cow per d for control (CON) or 56 g/cow per d for OG for 52 d on a commercial dairy. At 52 d of the study cows were randomly selected (n = 12) from both groups (6 OG and 6 CON) and housed in environmentally controlled rooms at the Agricultural Research Complex for 21 d at the University of Arizona. Cows were subjected to 7 d of thermoneutral (TN) conditions, 10 d of heat stress (HS), and 4 d of recovery (REC) under TN conditions. Feed intake, milk production, and milk composition were measured daily. Rectal temperatures (RT) and respiration rates (RR) were recorded 3 times per day (600, 1400, and 1800 h). Blood samples were taken on d 7 (TN), 8 (HS), 10 (HS), 17 (HS), and 18 (TN) during the Agricultural Research Complex segment. Cows in HS had higher RR and RT and water intake and lower dry matter intake and milk yield than these measures in TN. There was a treatment × environment interaction with cows fed OG having lower RR and RT and higher dry matter intake during peak thermal loads than CON. However, milk yield did not differ between groups. Cows fed OG had lower milk fat percent than CON (3.7 vs 4.3%) during HS. The SCC content of milk did not differ between treatment groups but rose in both groups during the REC phase following HS. Plasma insulin and plasma glucose levels were not different between groups. However, plasma insulin in both groups was lower during acute HS, then rose across the HS period, and was highest during the REC phase. Plasma cortisol levels were highest in all cows on the first day of HS (d 8) but were lower in cows fed OG compared with CON. However, plasma ACTH concentrations were elevated in OG-fed animals at all times samples were collected. Plasma ACTH was also elevated in cows fed both OG and CON during HS. Feeding OG reduced plasma cortisol during acute but not chronic HS and increased basal plasma ACTH, suggesting that OG treatment may alter the hypothalamic pituitary adrenal axis.     

Impacts of chronic and increasing lipopolysaccharide exposure on production and reproductive parameters in lactating Holstein dairy cows

Lipopolysaccharide (LPS) administration causes immunoactivation, which negatively affects production and fertility, but experimental exposure via an acute bolus is unlikely to resemble natural infections. Thus, the objectives were to characterize effects of chronic endotoxemia on production parameters and follicular development in estrous-synchronized lactating cows. Eleven Holstein cows (169 ± 20 d in milk; 681 ± 16 kg of body weight) were acclimated to their environmental surroundings for 3 d and then enrolled in 2 experimental periods (P). During P1 (3 d) cows consumed feed ad libitum and baseline samples were obtained. During P2 (7 d), cows were assigned to continuous infusion of either (1) saline-infused and pair-fed (CON-PF; 40 mL/h of saline i.v.; n = 5) or (2) LPS infused and ad libitum fed (LPS-AL; Escherichia coli O55:B5; 0.017, 0.020, 0.026, 0.036, 0.055, 0.088, and 0.148 μg/kg of body weight/h i.v. on d 1 to 7, respectively; n = 6). Controls were pair-fed to the LPS-AL group to eliminate confounding effects of dissimilar nutrient intake. Infusing LPS temporally caused mild hyperthermia on d 1 to 3 (+0.49°C) relative to baseline. Dry matter intake of LPS-AL cows decreased (28%) on d 1 of P2, then progressively returned to baseline. Relative to baseline, milk yield from LPS-AL cows was decreased on d 1 of P2 (12%). No treatment differences were observed in milk yield during P2. Follicular growth, dominant follicle size, serum progesterone (P4), and follicular P4 and 17β-estradiol concentrations were similar between treatments. Serum 17β-estradiol tended to increase (115%) and serum amyloid A and LPS-binding protein were increased (118 and 40%, respectively) in LPS-AL relative to CON-PF cows. Compared with CON-PF, neutrophils in LPS-AL cows were initially increased (45%), then gradually decreased. In contrast, monocytes were initially decreased (40%) and progressively increased with time in the LPS-AL cows. Hepatic mRNA abundance of cytochrome P450 family 2 subfamily C (CYP2C) or CYP3A was not affected by LPS, nor was there a treatment effect on toll-like receptor 4 or LBP; however, acyloxyacyl hydrolase and RELA subunit of nuclear factor kappa B tended to be increased in LPS-AL cows. These data suggest lactating dairy cows become tolerant to chronic and exponentially increasing LPS infusion in terms of production and reproductive parameters.     

Effects of pre- and postpartum dietary starch content on productivity,plasma energy metabolites,and serum inflammation indicators of dairy cows

The objective of this study was to evaluate the effects of the starch content of pre- and postpartum diets on productivity, plasma energy metabolites, and serum markers of inflammation of dairy cows during the calving transition period. Eighty-eight primiparous and multiparous cows were randomly assigned to pre- and postpartum dietary treatments balanced for parity and pretrial body condition score at d 28 ± 3 before expected calving date. Cows were fed either a control [Control; 14.0% starch, dry matter (DM) basis] or high-starch (High; 26.1% starch, DM basis) prepartum diet commencing 28 ± 3 d before expected calving date. Following calving, cows were fed either a high-fiber (HF; 33.8% neutral detergent fiber, 25.1% starch, DM basis) or high-starch (HS; 27.2% neutral detergent fiber, 32.8% starch, DM basis) postpartum diet for the first 20 ± 2 d following calving. Cows fed the High prepartum diet had greater DM intake (12.4 vs. 10.2 kg/d), plasma concentrations of insulin (1.72 vs. 14.2 ng/mL), glucose (68.1 vs. 65.0 mg/dL), and glucagon-like peptide-2 (0.41 vs. 0.32 ng/mL) before parturition, but increased plasma free fatty acid concentration (452 vs. 363 µEq/L) and milk fat yield (1.64 vs. 1.48 kg/d) after parturition. Cows fed the HS postpartum diet had lower plasma free fatty acid (372 vs. 442 µEq/L) and serum haptoglobin (0.46 vs. 0.70 mg/mL) concentrations over a 3-wk period after calving. In addition, there was a tendency for interaction between prepartum and postpartum diets for milk yield, where feeding the HS postpartum diet increased milk yield compared with the HF diet for cows fed the Control prepartum diet (40.8 vs. 37.9 kg/d) but not for cows fed the High prepartum diet. These results suggest that management efforts to minimize the change in diet fermentability during the calving transition by feeding the High prepartum diet, the HF postpartum diet, or both did not increase productivity of dairy cows but increased fat mobilization after calving. Our findings also suggest that feeding high-starch postpartum diets can decrease fat mobilization and serum indicators of systemic inflammation and increase milk production even with the transition from a low-starch prepartum diet.