Corrigendum to "Effects of feed additives on rumen function and bacterial and archaeal communities during a starch and fructose challenge" (J. Dairy Sci. 106:8787-8808).

Heifers (n = 40) were randomly allocated to 5 treatment groups: (1) control (no additives); (2) virginiamycin (VM; 200 mg/d); (3) monensin (MT; 200 mg/d) + tylosin (110 mg/d); (4) monensin (MLY; 220 mg/d) + live yeast (5.0 × 108 cfu/d); (5) sodium bicarbonate (BUF; 200 g/d) + magnesium oxide (30 g/d).

Effects of feed additives on rumen function and bacterial and archaeal communities during a starch and fructose challenge.

The objective of this study was to improve understandings of the rumen microbial ecosystem during ruminal acidosis and responses to feed additives to improve prudent use strategies for ruminal acidosis control. Rumen bacterial and archaeal community composition (BCC) and its associations with rumen fermentation measures were examined in Holstein heifers fed feed additives and challenged with starch and fructose. Heifers (n = 40) were randomly allocated to 5 treatment groups: (1) control (no additives); (2) virginiamycin (VM; 200 mg/d); (3) monensin (MT; 200 mg/d) + tylosin (110 mg/d); (4) monensin (MLY; 220 mg/d) + live yeast (5.0 × 1012 cfu/d); (5) sodium bicarbonate (BUF; 200 g/d) + magnesium oxide (30 g/d). Heifers were fed twice daily a 62% forage:38% concentrate total mixed ration at 1.25% of body weight (BW) dry matter (DM)/d for a 20-d adaptation period with their additive(s). Fructose (0.1% of BW/d) was added to the ration for the last 10 d of adaptation. On d 21 heifers were challenged once with a ration consisting of 1.0% of BW DM wheat and 0.2% of BW fructose plus their additive(s). A rumen sample was collected from each heifer via stomach tube weekly (d 0, 7, 14) and 5 times over a 3.6 h period at 5, 65, 115, 165, and 215 min after consumption of the challenge ration (d 21) and analyzed for pH, and ammonia, d- and l-lactate, volatile fatty acids (VFA), and histamine concentrations and total bacteria and archaea. The 16S rRNA gene spanning the V4 region was PCR amplified and sequenced. Alpha and ß diversity and associations of relative abundances of taxa with rumen fermentation measures were evaluated. Rumen BCC shifted among treatment groups in the adaptation period and across the challenge sampling period, indicating the feed additives had different modes of action. The monensin-containing treatment groups, MT and MLY often had similar relative abundances of rumen bacterial phyla and families. The MLY treatment group was characterized in the challenge period by increased relative abundances of the lactate utilizing genera Anaerovibrio and Megasphaera. The MLY treatment group also had increased diversity of ruminal bacteria which may provide resilience to changes in substrates. The control and BUF treatment groups were most similar in BCC. A redundancy analysis showed the MLY treatment group differed from all other treatment groups and concentrations of histamine and valerate in the rumen were associated with the most variation in the microbiota, 5.3% and 4.8%, respectively. It was evident from the taxa common to all treatment groups that cattle have a core microbiota. Functional redundancy of rumen bacteria which was reflected in the greater sensitivity for the rumen BCC than rumen fermentation measures likely provide resilience to changes in substrate. This functional redundancy of microbes in cattle suggests that there is no single optimal ruminal microbial population and no universally superior feed additive(s). In summary, differences in modes of action suggest the potential for more targeted and improved prudent use of feed additives with no single feed additive(s) providing an optimal BCC in all heifers.

Effect of prepartum source and amount of vitamin D supplementation on lactation performance of dairy cows.

The objectives of this experiment were to determine the effects of supplementing 25-hydroxyvitamin D3 (calcidiol, CAL) compared with vitamin D3 (cholecalciferol, CHOL) at 1 or 3 mg/d in late gestation on production outcomes of dairy cows. One hundred thirty-three parous and 44 nulliparous pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments (CAL1, CAL3, CHOL1, and CHOL3). Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d in gestation until parturition. The prepartum diet had a dietary cation-anion difference of -128 mEq/kg of dry matter. Production and disease were evaluated for the first 42 d in milk, and reproduction was evaluated to 300 d in milk. Incidence of postpartum diseases did not differ among treatments. Feeding CAL compared with CHOL increased yields of colostrum and colostrum fat, protein, and total solids, resulting in an increased amount of net energy for lactation secreted as colostrum (CHOL = 7.0 vs. CAL = 9.0 ± 0.7 Mcal). An interaction between source and amount was observed for milk yield: CAL3 increased milk yield compared with CHOL3 (CHOL3 = 34.1 vs. CAL3 = 38.7 ± 1.4 kg/d) but milk yield did not differ between CAL1 and CHOL1 (CHOL1 = 36.9 vs. CAL1 = 36.4 ± 1.4 kg/d). Concentrations of serum calcidiol on day of calving and average serum Ca from d 2 to 11 postpartum were positively associated with milk yield in the first 42 d in milk. Interactions between source and amount of vitamin D were also observed for pregnancy after first AI: the percentage of cows receiving CHOL1 and CAL3 that became pregnant was smaller than that of cows receiving CHOL3 and CAL1. However, pregnancy per AI and pregnancy by 300 d in milk did not differ among treatments. Overall, CAL3 increased milk yield compared with CHOL3, whereas in cows fed 1 mg/d (CAL1 and CHOL1), the source of vitamin D generally had no effect. The effect of CAL3 may be explained in part by serum CAL concentrations and postpartum serum Ca, which were associated with milk yield.

Effect of vitamin D source and amount on vitamin D status and response to endotoxin challenge.

The objectives were to test the effects of dietary vitamin D3 [cholecalciferol (CHOL)] compared with 25-hydroxyvitamin D3 [calcidiol (CAL)] on vitamin D status and response to an endotoxin challenge. Forty-five Holstein bull calves (5 ± 2 d of age) were blocked into weekly cohorts, fed a basal diet that provided 0.25 µg/kg body weight (BW) CHOL, and assigned randomly to 1 of 5 treatments: control [(CON) no additional vitamin D], 1.5 µg/kg BW CHOL (CHOL1.5), 3 µg/kg BW CHOL (CHOL3), 1.5 µg/kg BW CAL (CAL1.5), or 3 µg/kg BW CAL (CAL3). Calves were fed milk replacer until weaning at 56 d of age and had ad libitum access to water and starter grain throughout the experiment. Treatments were added daily to the diet of milk replacer until weaning and starter grain after weaning. Measures of growth, dry matter intake, and serum concentrations of vitamin D, Ca, Mg, and P were collected from 0 to 91 d of the experiment. At 91 d of the experiment, calves received an intravenous injection of 0.1 µg/kg BW lipopolysaccharide (LPS). Clinical and physiological responses were measured from 0 to 72 h relative to LPS injection. Data were analyzed with mixed models that included fixed effects of treatment and time, and random effect of block. Orthogonal contrasts evaluated the effects of (1) source (CAL vs. CHOL), (2) dose (1.5 vs. 3.0 µg/kg BW), (3) interaction between source and dose, and (4) supplementation (CON vs. all other treatments) of vitamin D. From 21 to 91 d of the experiment, mean BW of supplemented calves was less compared with CON calves, but the effect was predominantly a result of the CHOL calves, which tended to weigh less than the CAL calves. Supplementing vitamin D increased concentrations of 25-hydroxyvitamin D in serum compared with CON, but the increment from increasing the dose from 1.5 to 3.0 µg/kg BW was greater for CAL compared with CHOL (CON = 18.9, CHOL = 24.7 and 29.6, CAL = 35.6 and 65.7 ± 3.2 ng/mL, respectively). Feeding CAL also increased serum Ca and P compared with CHOL. An interaction between source and dose of treatment was observed for rectal temperature and derivatives of reactive metabolites after LPS challenge because calves receiving CHOL3 and CAL1.5 had lower rectal temperatures and plasma derivatives of reactive metabolites compared with calves receiving CHOL1.5 and CAL3. Supplementing vitamin D increased plasma P concentrations post-LPS challenge compared with CON, but plasma concentrations of Ca, Mg, fatty acids, glucose, ß-hydroxybutyrate, haptoglobin, tumor necrosis factor-α, and antioxidant potential did not differ among treatments post-LPS challenge. Last, supplementing vitamin D increased granulocytes as a percentage of blood leukocytes post-LPS challenge compared with CON. Supplementing CAL as a source of vitamin D to dairy calves was more effective at increasing serum 25-hydroxyvitamin D, Ca, and P concentrations compared with feeding CHOL. Supplemental source and dose of vitamin D also influenced responses to the LPS challenge.

Effect of source and amount of vitamin D on serum concentrations and retention of calcium, magnesium, and phosphorus in dairy cows.

The objectives of the experiment were to determine the effects of supplementing 2 amounts of 25-hydroxyvitamin D3 (calcidiol; CAL) compared with equal amounts of vitamin D3 (cholecalciferol; CHOL) on serum concentrations, absorptions, and retentions of Ca, Mg, and P in periparturient dairy cows. One hundred seventy-seven (133 parous and 44 nulliparous) pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit for energy-corrected milk yield (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments. Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d gestation until parturition. The prepartum diet had a dietary cation-anion difference of -128 mEq/kg of dry matter. All cows were fed a common postpartum diet containing 46 µg of vitamin D3/kg of dry matter without further supplementation of treatments. Concentrations of vitamin D metabolites, Ca, Mg, and P in serum were measured pre- and postpartum, in addition to total-tract digestibility and urinary excretion of Ca, Mg, and P in the prepartum period. Feeding 3 mg compared with 1 mg of CAL increased serum 25-hydroxyvitamin D3 (CAL1 = 94 vs. CAL3 = 173 ± 3 ng/mL). In comparison, the increment in serum 25-hydroxyvitamin D3 from feeding 3 mg compared with 1 mg of CHOL was small (CHOL1 = 58 vs. CHOL3 = 64 ± 3 ng/mL). Feeding CAL increased prepartum concentration of P in serum compared with CHOL (CHOL = 1.87 vs. CAL = 2.01 ± 0.02 mM), regardless of the amount fed, but neither source nor amount affected prepartum Ca or Mg in serum. Feeding CAL increased serum Ca and P for the first 11 d postpartum compared with CHOL (CHOL = 2.12 vs. CAL = 2.16 ± 0.01 mM serum Ca; CHOL = 1.70 vs. CAL = 1.78 ± 0.02 mM serum P) but the amount of vitamin D did not affect postpartum concentrations of Ca, Mg, and P in serum. Feeding CAL increased prepartum apparent digestibility of Ca compared with CHOL (CHOL = 26.6 vs. CAL = 33.5 ± 2.8%) but treatments did not affect Ca retention prepartum. Neither source nor amount of vitamin D affected Mg and P apparent digestibility, but CAL decreased the concentration of P excreted in urine during the prepartum period (CHOL = 1.8 vs. CAL = 0.8 ± 0.3 g/d). Calcidiol tended to increase the amount of Ca secreted in colostrum (CHOL = 9.1 vs. CAL = 11.2 ± 0.9 g/d) and Ca excreted in urine postpartum (CHOL = 0.4 vs. CAL = 0.6 ± 0.1 g/d) compared with CHOL. Collectively, feeding CAL at 1 or 3 mg/d compared with CHOL in the last 24 d of gestation is an effective way to increase periparturient serum P concentration and postpartum serum Ca of dairy cows fed a prepartum diet with negative DCAD.

Epigallocatechin-3-gallate protected vanadium-induced eggshell depigmentation via P38MAPK-Nrf2/HO-1 signaling pathway in laying hens.

It has been demonstrated that tea polyphenol (TP) epigallocatechin-3-gallate (EGCG) can confer protection against vanadium (V) toxicity in laying hens; however, our understanding of the molecular mechanisms beyond this effect are still limited. In this study, 360 hens were randomly assigned to the 3 groups to study whether the potential mechanism P38MAPK-Nrf2/HO-1 signaling pathway is involved in the protective effect of EGCG on eggshell pigmentation in vanadium challenged laying hens. Treatments included a control group, a 10 mg/kg V (V10), and a V10 plus 130 mg/kg of EGCG group (V10+EGCG130). Both eggshell color and protoporphyrin IX were decreased in the V10 group compared with the control diet, while EGCG130 treatment partially improved shell color and protoporphyrin IX (P < 0.05). The V10 exposure induced higher cell apoptosis rate and oxidative stress in birds as evidenced by the histological apoptosis status, decreased uterine glutathione-S transferase (GST) and high abundance of malondialdehyde (MDA) compared with the control group, whereas EGCG130 markedly alleviated oxidative stress via reducing MDA generation (P < 0.05). Dietary vanadium reduced ferrochelatase, NF-E2-related factor 2 (Nrf2), and heme oxygenase (HO-1) mRNA expression, while EGCG up-regulated Nrf2 and HO-1 expression (P < 0.05). Protein levels of Nrf2, HO-1 and phospho-p38 (P-P38) MAPK were reduced in V10 group, while dietary supplementation with 130 mg/kg EGCG markedly increased Nrf2, HO-1 and P-P38 MAPK protein levels in the uterus compared with the V10 group (P < 0.01). In conclusion, EGCG improved eggshell color and antioxidant system in V10-challenged hens, which seems to be associated with P38MAPK-Nrf2/HO-1 signaling pathway.

Associations between bone and energy metabolism in cows fed diets differing in level of dietary cation-anion difference and supplemented with cholecalciferol or calcidiol.

Bone-derived hormones play an important role in metabolism. This study examined the hypothesis that interactions between bone and energy metabolism, particularly those involving osteocalcin, are present in dairy cattle and have feedback mechanisms over time. Associations between metabolites in blood were examined in 32 Holstein cows blocked by parity and milk yield and randomly allocated to diets containing either 0.27 mg/kg dry matter (DM) calcidiol or cholecalciferol for an anticipated intake of 3 mg/d (120,000 IU/d) at 11 kg of DM, and positive (+130 mEq/kg DM) or negative (-130 mEq/kg DM) dietary cation-anion difference (DCAD) from 252 d of gestation to calving. Blood was sampled every 3 d, from 9 d prepartum to 30 d postpartum, and plasma concentrations of vitamin D3, 25-hydroxyvitamin D3, adiponectin, C-telopeptide of type 1 collagen (CTX1), glucose, insulin-like growth factor 1 (IGF1), insulin, undercarboxylated osteocalcin (uOC), and carboxylated osteocalcin (cOC) were determined. Feeding calcidiol compared with cholecalciferol increased plasma concentrations of 25-hydroxyvitamin D3 pre- (264.2 ± 8.0 vs. 61.3 ± 8.0 ng/mL) and postpartum (170.8 ± 6.2 vs. 51.3 ± 6.2 ng/mL) but decreased concentrations of vitamin D3 pre- (1.2 ± 0.6 vs. 14.5 ± 0.6 ng/mL) and postpartum (1.9 ± 0.4 vs. 3.2 ± 0.6 ng/mL). Prepartum, cows fed the negative DCAD diet had reduced concentrations of vitamin D3 and glucose compared with cows fed a positive DCAD. The combination of negative DCAD and cholecalciferol reduced IGF1 concentrations prepartum. The DCAD treatment had no effect on postpartum concentrations of metabolites. Nulliparous cows had increased concentrations of OC, CTX1, IGF1, glucose, and insulin compared with parous cows. Time series analysis identified associations between metabolites on the same day and over 3-d lags up to ±9 d that suggest feedback between 25-hydroxyvitamin D3 and vitamin D3 in the negative lags, indicating that 25-hydroxyvitamin D3 may exert feedback on vitamin D3 but not vice versa. We found evidence of a feedback mechanism between vitamin D3 and IGF1, with positive effect size (ES) on the same day and 3 d later, and negative ES 9 d later, that was more evident in cholecalciferol-fed cows. This suggests an important role of IGF1 in integrating bone metabolism with energy and protein metabolic pathways. Evidence of feedback was found between uOC and particularly cOC with IGF1, with positive ES on the same day but negative ES 6 d before and 6 d after. An association between uOC or cOC and IGF1 has not been previously identified in cattle and suggests that both uOC and cOC may have marked biological activity. Associations between OC and insulin identified in mice were not observed herein, although associations between OC and glucose were similar to those between IGF1 and glucose, supporting associations between glucose, OC, and IGF1. We provide further statistical evidence of crosstalk between vitamin D compounds, bone hormones, and energy metabolism in cattle. In particular, associations between uOC or cOC and IGF1 may provide links between prepartum diets and observations of prolonged increases in milk production and allow better control of peripartum metabolism.

Effects of nutrition on the fertility of lactating dairy cattle.

This meta-analysis of 39 experiments containing 118 treatments explored the effects of diet interventions in early lactation on the proportion of dairy cows pregnant to artificial insemination (AI; pregnancy to AI) and on calving to pregnancy interval. It also identified factors that may explain variation in these responses. The objectives were to identify effects of diet on reproduction, rather than differences between specific dietary interventions. The examination of calving to pregnancy interval used the more traditional method of analyzing differences between a treatment and the reference treatment used for comparison within a given experiment. The systematic review identified fewer experiments (n = 39) than had been expected. Four different multivariable models including the random effect of experiment were used to examine the effects of CPM-Dairy (version 3.08) estimated diet and production variables on proportion pregnant to AI. These models examined (1) output of products, (2) balance or duodenal availability of nutrients, (3) intake of nutrients, or (4) percentage of nutrients in the diet. The multivariable models identified positive associations between estimated increased fatty acid intake [incidence rate ratio (IRR) = 1.0003 ± 0.0001g/d; ±standard error], starch intake (IRR = 1.061 ± 0.029 kg/d), metabolizable energy balance (IRR = 1.004 ± 0.002 MJ/d), and duodenal C14:0 (IRR = 1.008 ± 0.004 g/d) availability with the proportion of cows pregnant to AI, whereas rapidly fermentable sugar intake (IRR = 0.813 ± 0.054 kg/d), percentage of sugar in the diet (IRR = 0.960 ± 0.015%), and milk protein yield (IRR 0.922 ± 0.022 g/100 g per day) were associated with a reduced proportion of cows pregnant to AI. There was no multivariable model developed to assess variables associated with calving to pregnancy interval but, univariably, increased metabolizable energy balance was associated with a shorter calving to pregnancy interval whereas increased milk production was associated with longer time to pregnancy. Increased intake of some AA, particularly threonine and lysine, were associated with a longer calving to pregnancy interval. It is clear nutritional management around calving can influence reproductive success. The importance of dietary fats and increased energy and protein balances in early lactation for improved fertility outcomes is supported and suggests that starch and sugars may have different effects on the proportion of cows that are pregnant to AI. This work also highlighted a need for further focused field studies exploring the roles of specific fatty acids, AA, phosphorus, and carbohydrates on reproduction.

Effects of a short-term supranutritional selenium supplementation on redox balance, physiology and insulin-related metabolism in heat-stressed pigs.

Heat stress (HS) disrupts redox balance and insulin-related metabolism. Supplementation with supranutritional amounts of selenium (Se) may enhance glutathione peroxidase (GPX) activity and reduce oxidative stress, but may trigger insulin resistance. Therefore, the aim of this experiment was to investigate the effects of a short-term high Se supplementation on physiology, oxidative stress and insulin-related metabolism in heat-stressed pigs. Twenty-four gilts were fed either a control (0.20 ppm Se) or a high Se (1.0 ppm Se yeast, HiSe) diet for 2 weeks. Pigs were then housed in thermoneutral (20°C) or HS (35°C) conditions for 8 days. Blood samples were collected to study blood Se and oxidative stress markers. An oral glucose tolerance test (OGTT) was conducted on day 8 of thermal exposure. The HS conditions increased rectal temperature and respiration rate (both p < .001). The HiSe diet increased blood Se by 12% (p < .05) and ameliorated the increase in rectal temperature (p < .05). Heat stress increased oxidative stress as evidenced by a 48% increase in plasma advanced oxidized protein products (AOPPs; p < .05), which may be associated with the reductions in plasma biological antioxidant potential (BAP) and erythrocyte GPX activity (both p < .05). The HiSe diet did not alleviate the reduction in plasma BAP or increase in AOPPs observed during HS, although it tended to increase erythrocyte GPX activity by 13% (p = .068). Without affecting insulin, HS attenuated lipid mobilization, as evidenced by a lower fasting NEFA concentration (p < .05), which was not mitigated by the HiSe diet. The HiSe diet increased insulin AUC, suggesting it potentiated insulin resistance, although this only occurred under TN conditions (p = .066). In summary, HS induced oxidative stress and attenuated lipid mobilization in pigs. The short-term supranutritional Se supplementation alleviated hyperthermia, but did not protect against oxidative stress in heat-stressed pigs.

Effects of prepartum dietary cation-anion difference and source of vitamin D in dairy cows: Vitamin D, mineral, and bone metabolism.

Pregnant Holstein cows, 28 nulliparous and 51 parous, were blocked by parity and milk yield and randomly allocated to receive diets that differed in dietary cation-anion difference (DCAD), +130 or -130 mEq/kg, and supplemented with either calcidiol or cholecalciferol at 3 mg/11 kg of dry matter from 255 d of gestation until parturition. Blood was sampled thrice weekly prepartum, and on d 0, 1, 2, 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30 postpartum to evaluate effects of the diets on vitamin D, mineral and bone metabolism, and acid-base status. Blood pH and concentrations of minerals, vitamin D metabolites, and bone-related hormones were determined, as were mineral concentrations and losses in urine and colostrum. Supplementing with calcidiol increased plasma concentrations of 25-hydroxyvitamin D3, 3-epi 25-hydroxyvitamin D3, 25-hydroxyvitamin D2, 1,25-dihydroxyvitamin D3, and 24,25-dihydroxyvitamin D3 compared with supplementing with cholecalciferol. Cows fed the diet with negative DCAD had lesser concentrations of vitamin D metabolites before and after calving than cows fed the diet with positive DCAD, except for 25-hydroxyvitamin D2. Feeding the diet with negative DCAD induced a compensated metabolic acidosis that attenuated the decline in blood ionized Ca (iCa) and serum total Ca (tCa) around calving, particularly in parous cows, whereas cows fed the diet with positive DCAD and supplemented with calcidiol had the greatest 1,25-dihydroxyvitamin D3 concentrations and the lowest iCa and tCa concentrations on d 1 and 2 postpartum. The acidogenic diet or calcidiol markedly increased urinary losses of tCa and tMg, and feeding calcidiol tended to increase colostrum yield and increased losses of tCa and tMg in colostrum. Cows fed the diet with negative DCAD had increased concentrations of serotonin and C-terminal telopeptide of type 1 collagen prepartum compared with cows fed the diet with positive DCAD. Concentrations of undercarboxylated and carboxylated osteocalcin and those of adiponectin did not differ with treatment. These results provide evidence that dietary manipulations can induce metabolic adaptations that improve mineral homeostasis with the onset of lactation that might explain some of the improvements observed in health and production when cows are fed diets with negative DCAD or supplemented with calcidiol.

Effects of chromium supplementation on physiology, feed intake, and insulin related metabolism in growing pigs subjected to heat stress.

Improving insulin sensitivity may reduce impacts of heat stress (HS) in pigs by facilitating heat dissipation. Chromium (Cr) has been reported to improve insulin sensitivity in pigs. Therefore, the aim of this experiment was to investigate whether Cr supplementation can mitigate HS in growing pigs. Thirty-six gilts were randomly assigned to 2 diets containing 0 (control) or 400 ppb Cr. After 14 d the supplemented pigs were allocated to either 8 d thermoneutral (20°C constant; TN) or cyclic HS (35°C, 0900 h to 1700 h) conditions and continued their respective diet (n = 9 per group). Growth performance was recorded during the 14-d supplementation period. The physiological responses to HS were monitored by measuring respiration rate, rectal temperature, blood gas chemistry, and feed intake during thermal exposure. Kinetics of plasma glucose, insulin and NEFA were studied by intravenous glucose tolerance test (IVGTT) on d 8 of thermal treatment. Results showed Cr alleviated the HS-increased rectal temperature (P < 0.05) and respiration rate (P < 0.01) at 1300 h and 1600 h during thermal exposure. However, Cr did not mitigate the reduction in average daily feed intake which was reduced by 35% during HS or the HS-induced respiratory alkalosis. Chromium tended to increase average daily gain (0.86 vs. 0.95 kg, P = 0.070) during the 14-d supplementation under TN conditions before thermal exposure, which might be associated with the potential of Cr in improving overall insulin sensitivity, as evidenced by a reduced insulin resistance index calculated by Homeostatic Model Assessment (HOMA-IR; 0.65 vs. 0.51, P = 0.013) and a tendency of reduced fasting plasma insulin concentration (1.97 vs. 1.67 µU/mL, P = 0.094). Heat stress decreased the acute insulin releasing rate (P = 0.012) and consequently slowed glucose clearance rate (P = 0.035) during IVGTT. Besides, HS enlarged the values of area under the curve of NEFA during IVGTT (P < 0.01), indicating a reduced lipid mobilization. In conclusion, HS reduced insulin response to IVGTT. Chromium supplementation exhibited a potential in improving insulin sensitivity and mitigating HS symptoms in growing pigs.

Risk factors in equine transport-related health problems: A survey of the Australian equine industry.

BACKGROUND: Transportation can affect equine health and is a potential source of economic loss to the industry. OBJECTIVES: To identify journey (duration, vehicle, commercial or noncommercial) and horse (sex, age, breed, use, amateur or professional status) characteristics associated with the development of transport-related health problems in horses. STUDY DESIGN: Cross-sectional online survey. METHODS: An online survey was conducted targeting amateur and professional participants in the Australian equine industry; eligible respondents were required to organise horse movements at least monthly. Respondents provided details of the last case of a transport-related health problem that had affected their horse(s). Associations between type of health problem, journey and horse characteristics were examined with multivariable multinomial regression analysis. RESULTS: Based on 214 responses, health problems were classified as injuries, muscular problems, heat stroke, gastrointestinal and respiratory problems, and death or euthanasia. Respiratory problems were reported most frequently (33.7%), followed by gastrointestinal problems (23.8%) and traumatic injuries (16.3%). The type of health problem was associated with journey duration (P<0.001) and horse breed (P = 0.001). Injuries were more likely to occur on short journeys, whereas more severe illnesses (gastrointestinal and respiratory problems, and death or euthanasia) were more likely to occur on long journeys. Using Standardbreds as the reference group, Thoroughbreds, Arabians and Warmbloods were more likely to experience a severe illness than an injury. MAIN LIMITATIONS: Self-selected participation in the study and the self-reported nature of transport-related problems. CONCLUSIONS: Horses undertaking journeys of longer than 24 h are at greater risk for the development of severe disease or death. Further studies on long-haul transportation effects are required to safeguard the welfare of horses moved over long distances.

Survey of horse transportation in Australia: issues and practices.

OBJECTIVE: To survey amateur and professional participants on equine transportation management, practices and outcomes in Australia. METHODS: An online survey targeting people who organised horse movements at least monthly was made available to a broad cross-section of amateur and professional equine associations. Respondents were invited to provide demographic details and information relating to their routine transportation management practices and their experiences of issues relating to the transportation of horses. RESULTS: Of 797 usable responses involving approximately 17,000 horses and 313,000 individual horse transport events, transport-related behavioural problems were reported by 38% of respondents, particularly at loading. Transport-related health problems had been experienced during or after transportation by horses in the care of 67% of respondents. The most common problems reported were traumatic injuries (45.0%), diarrhoea (20.0%), muscular problems (13.0%), respiratory problems (12.3%), overheating (10.5%) and colic (10.3%). In the 2 years reviewed in the survey, 9.4% of participants reported at least one case of transport-associated pneumonia and 35 horses had died, most commonly from fractures, colic or pneumonia. Although respondents identifying as amateurs transported horses less frequently and over shorter distances, the incidence of transport-related problems was similar between amateurs and professionals. Respondents reported specific precautions before, during and after transportation, although management was often not compliant with the Australian Code of horse transportation. CONCLUSIONS: Responses indicated that there remains a substantial risk of adverse welfare and health outcomes for horses transported in Australia and management practices reported may not be compliant with current recommendations for transportation.

High dietary selenium and vitamin E supplementation ameliorates the impacts of heat load on oxidative status and acid-base balance in sheep.

The objective of this study was to determine the efficacy of supranutritional dietary selenium and vitamin E (Vit E) to ameliorate the effect of heat stress (HS) on oxidative status and acid-base balance in sheep. Thirty-two Merino × Poll Dorset ewes were acclimated to indoor individual pen feeding of a pelleted control diet (0.24 g Se and 10 IU of Vit E/kg DM) for 1 wk. Sheep were then moved to metabolism cages in climatic chambers and randomly allocated to a 2 × 2 × 2 factorial design with the respective factors being dietary Se (0.24 and 1.20 mg/kg DM as Sel-Plex; Alltech, Australia), Vit E (10 and 100 IU/kg DM), and temperature for 2 wk. After 1 wk of acclimation in metabolic cages, 1 climatic chamber continued on thermoneutral (TN) conditions (18°C to 21°C and 40% to 50% relative humidity [RH]), and the other one was set to HS conditions (28°C to 40°C and 30% to 40% RH) for 1 wk. The sheep were then returned to individual pens and fed the control diet for 1 wk before being returned to the same diet as in the first period but a reversed thermal treatment for a further 2 wk. Physiological parameters were recorded 3 times daily, and blood samples were collected on d 1 and 7 of thermal treatment. Average respiration rate and rectal temperature of sheep were increased (P < 0.001) during HS; however, combined supranutritional supplementation of Se and Vit E reversed the effects of HS. Sheep given the high Se and high Vit E diet had a lower respiration rate (191 vs. 232 breaths/min; P = 0.012) and rectal temperature (40.33°C vs. 40.58°C; P = 0.039) under peak HS (1700 h) compared with those fed the low Se and low Vit E diet. Plasma reactive oxygen metabolites concentrations were reduced (P = 0.048) by 20%, whereas biological antioxidant potential was increased (P = 0.17) by 10% in sheep fed the high Se and high Vit E diet compared with those fed the low Se and low Vit E diet. Blood pH was elevated (P = 0.007) and bicarbonate was reduced (P = 0.049) under HS, and again, these effects were ameliorated by the high Se and high Vit E diet. Both white blood cell glutathione peroxidase gene expression and red blood cell lysate glutathione peroxidase activity were increased in sheep fed the high Se and high Vit E diet. These data suggest that supranutritional dietary Se or Vit E can reduce some of the negative effects of HS. However, the synergism between the 2 antioxidants improves their potential to ameliorate the impacts of HS in sheep.

Effects of dietary fat on fertility of dairy cattle: A meta-analysis and meta-regression.

Evidence is increasing of positive effects of feeding fats during transition on fertility and the adaptation to lactation. This study used meta-analytic methods to explore the effects of including fats in the transition diet on the risk of pregnancy to service (proportion pregnant) and calving to pregnancy interval. Meta-analysis was used to integrate smaller studies and increase the statistical power over that of any single study and explore new hypotheses. We explored the effect of fats and diet composition on fertility using meta-regression methods. Relatively few highly controlled studies are available providing detailed descriptions of the diets used that examined interactions between fat nutrition and reproductive outcomes. Only 17 studies containing 26 comparisons were suitable for inclusion in statistical evaluations. Reproductive variables evaluated were risk of pregnancy (proportion pregnant), primarily to first service, and calving to pregnancy interval. Production variables examined were milk yield, milk composition, and body weight. The sources of heterogeneity in these studies were also explored. A 27% overall increase in pregnancy to service was observed (relative risk=1.27; 95% confidence interval Knapp Hartung 1.09 to 1.45), and results were relatively consistent (I(2)=19.9%). A strong indication of a reduction in calving to pregnancy interval was also identified, which was consistent across studies (I(2)=0.0%), supporting a conclusion that, overall, the inclusion of fats does improve fertility. Further exploration of the factors contributing to proportion pregnant using bivariate meta-regression identified variables that reflected changes in diet composition or animal response resulting from inclusion of the fat interventions in the experimental diets fed. Increased fermentable neutral detergent fiber and soluble fiber intakes increased the proportion pregnant, whereas increased milk yield of the treatment group decreased this measure. Unexpectedly, the estimated energy costs of urea production also had a positive association with proportion pregnant. The limited number of suitable studies for the analysis highlights the need for more work to improve understanding of the critical nutritional factors affecting fertility. These factors include specific fatty acids in dietary interventions that contribute to increasing fertility of cows in dairy production systems.

Changes in milk oxidative stress biomarkers in lactating dairy cows with ovulatory and an-ovulatory oestrous cycles.

This study was conducted to evaluate changes in milk profiles of oxidative stress (OS) biomarkers in dairy cows with ovulatory and an-ovulatory oestrous cycles. Thirty healthy, cycling Holstein cows averaging 60±17 days in milk, and producing 33±6kg of milk per day (the week before commencing the study) were enrolled in this study. Composite milk samples were collected thrice weekly and assayed for the following OS biomarkers: lipoperoxides (LPO), biological advanced potential, superoxide dismutase (SOD), advanced oxidation protein products (AOPP), ceruloplasmin, glutathione (GSH), ß-carotene and glutathione peroxidase (GSH-Px). Milk samples were also tested for fat and protein composition and the fat:protein ratio (FPR) was categorized as low (≤1.31), medium (1.32-1.56) and high (>1.57) to evaluate their main effect and the interaction effect of FPR and the week of study on OS using linear mixed models with cow identification being a random factor. Cows with ovulatory oestrous cycles (n=20) presented significantly greater SOD levels than cows that did not ovulate ((n=10; P<0.05). On the other hand, LPO, GSH-Px and GSH concentrations were lower in ovulated cows compared to the an-ovulated cows (P<0.05). The highest level of LPO and AOPP were noted at prooestrus phase while ß-carotene presented the lowest value at that phase of oestrous cycle. It could be postulated that the elevated level of milk SOD and the observed lower level of LPO, GSH-Px and GSH in ovulating cows may be an essential event preceding the ovulatory response.

Evaluation of infrared thermography body temperature and collar-mounted accelerometer and acoustic technology for predicting time of ovulation of cows in a pasture-based system.

This study was conducted to test the hypothesis that the specificity of infrared thermography (IRT) in detecting cows about to ovulate could be improved using different body parts that are less likely to be contaminated by fecal matter. In addition, the combined activity and rumination data captured by accelerometers were evaluated to provide a more accurate indication of ovulation than the activity and rumination data alone. Thermal images of 30 cows were captured for different body areas (eye, ear, muzzle, and vulva) twice daily after AM and PM milking sessions during the entire experimental period. Milk progesterone data and insemination records were used to determine the date of ovulation. Cows were fitted with SCR heat and rumination long-distance tags (SCR HR LD) for 1 month. Activity- and rumination-based estrus alerts were initially identified using default threshold values set by the manufacturer; however, a range of thresholds was also created and tested for both activity and rumination to determine the potential for higher levels of accuracy of ovulation detection. Visual assessment of mounting indicators resulted in 75% sensitivity (Se), 100% specificity (Sp), and 100% positive predictive value (PPV). Overall, IRT showed poor performance for detecting cows about to ovulate. Vulval temperature resulted in the greatest (80%) Sp but the poorest (21%) Se compared with the IRT temperatures of other body areas. The SCR HR LD tags default threshold value resulted in 78% Se, 57% Sp, and 70% PPV. Lowering the activity threshold from the default value improved the sensitivity but created a large number of false positives, which resulted in a decrease in specificity. Lowering the activity threshold to 20 resulted in a detection performance of 80% Se, 94% Sp, and 67% PPV, whereas the rumination levels achieved 35% Se, 69% Sp, and 14% PPV. The area under the curve for the activity level, rumination level, and the combined measures of activity and rumination levels were 0.82, 0.54, and 0.75, respectively. Alerts generated by SCR HR LD tags based on a lower activity threshold level had high sensitivity and may be able to detect a high proportion of cows in ovulatory periods in pasture-based system; however, the specificities and positive predictive value were lower than the visual assessment of mounting indicators.

Dietary antioxidants at supranutritional doses modulate skeletal muscle heat shock protein and inflammatory gene expression in sheep exposed to heat stress.

The objective of this study was to investigate the effects of chronic heat (thermal) stress and dietary antioxidant supplementation on the expression of heat shock proteins and inflammatory genes in the skeletal muscle of sheep. Twenty-four Merino × Poll Dorset crossbred ewes were allocated to either a control (10 IU vitamin E and 0.24 mg Se/kg DM) or high-antioxidant (VitE+Se; 100 IU vitamin E and 1.20 mg Se/kg DM) diet and were exposed to 2 thermal (temperature) treatments (thermoneutral [TN]: 18°C-21°C and 26%-30% relative humidity; heat stress [HS]: 28°C-40°C and 40%-50% relative humidity) for 1 wk. Physiological parameters were recorded daily, and muscle biopsies were conducted at the end of thermal treatments. Total RNA was extracted from muscle samples and reverse transcribed to cDNA for real-time PCR analysis. Respiration rates and rectal temperature were increased in response to HS (84.2 vs. 161 breaths per minute and 39.52°C vs. 40.06°C for TN and HS conditions, respectively; P < 0.001). There were interactions between dietary and thermal treatments, indicating that dietary antioxidant supplementation reduced respiration rate (P = 0.097) and rectal temperature (P = 0.086) of sheep during HS but not TN conditions. Skeletal muscle heat shock transcription factor 1 (HSF1) mRNA abundance was increased by HS (1.3-fold; P < 0.050) but was not changed (P = 0.77) by dietary antioxidant supplementation. The expression of skeletal muscle heat shock protein 70 (HSP70) mRNA was increased (P < 0.001) 3.5-fold by HS and tended (P = 0.08) to be increased by dietary antioxidant supplementation. Although there were no main effects of diet (P = 0.42) or HS (P = 0.47) on skeletal muscle HSP90 mRNA expression, there was an interaction (P = 0.040) such that HSP90 mRNA expression was increased (P = 0.010) in antioxidant-supplemented sheep under HS compared to TN conditions. Skeletal muscle nuclear factor kappa B (NF-κB) and tissue necrosis factor α (TNF-α) mRNA abundances were increased by exposure to heat (5.2-fold, P = 0.005 for NF-κB; 5.7-fold, P = 0.013 for TNF-α) ,but there was no main effect (P > 0.05) of dietary antioxidant supplementation. There was, however, an interaction between thermal and dietary treatments such that dietary antioxidant supplementation ameliorated the effect of HS on NF-κB and TNF-α mRNA expression. Taken together, these results indicate that high dietary antioxidants modulate skeletal muscle expression of heat shock proteins, proinflammatory cytokine, and NF-κB transcription, which may protect against HS in sheep.

Plasma oxidative stress biomarkers and progesterone profiles in a dairy cow diagnosed with an ovarian follicular cyst.

This study was conducted to examine the oxidative stress biomarkers in a cow diagnosed with a follicular cyst in her left ovary. Progesterone (P4) and plasma oxidative stress status was measured in 13 Holstein cows after synchronization of oestrus with controlled internal drug release (CIDR) and prostaglandinF2α (PGF2α) protocol. The presence and size of ovarian structures were monitored by transrectal ultrasound at 4 hourly intervals. Of the 13 cows, 12 were monitored until ovulation was detected and recorded, whereas one cow failed to ovulate and developed a follicular cyst. Oxidative stress biomarkers; reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP), oxidative stress index (OSI), glutathione (GSH), ceruloplasmin and advanced oxidation protein products (AOPP) were measured in the cystic cow and compared to those of the 12 ovulated cows and are referred to as higher or lower if they are outside the mean ± standard error of mean of those of ovulated cows. The cystic cow had lower ROMs and OSI between 36 and 84 h after PGF2α injection and at 9 h, from 36 to 60 h after PGF2α injection respectively. On the other hand, antioxidant (BAP and GSH) was higher in the cystic cow compared to her ovulated herd mates. The observed imbalance between oxidant and antioxidant might have disrupted the physiological events for ovulation to occur, leading to cystic ovarian disease.