Ovarian follicular activity in lactating Holstein cows supplemented with monensin

The objective of this study was to determine effects of monensin on ovarian follicular development and reproductive performance in postpartum dairy cows. Forty-eight multiparous Holstein cows were randomly assigned to receive either a control total mixed ration (n = 24) or the same diet plus 22 mg of monensin/kg (n = 24) from 21 d before anticipated calving until cows were either confirmed pregnant or were >180 d postpartum. Monensin had no effect on development of the first dominant follicle postpartum or the numbers of class 1 (3 to 5 mm), 2 (6 to 9 mm), or 3 (10 to 15 mm) follicles. Control cows had more class 4 (>15 mm) follicles at 10 to 13 d postpartum than cows in the monensin group. The first dominant follicle postpartum ovulated, regressed, or became cystic unrelated to differences between diets. However, the first ovulation postpartum occurred earlier in monensin-fed cows than in the control group (27.2 +/- 2.1 d vs. 32.4 +/- 1.5 d), with no dietary effects on the diameter of the ovulating follicle. Similarly, treatments did not differ in the proportion of cows with 2 or 3 waves of ovarian follicular development per cycle, nor in the number of follicles of all classes during the breeding period. Times of ovulation following treatment with prostaglandin F2alpha were not different between dietary groups. Pregnancy rates after timed artificial insemination were similar between diets. Supplementation with monensin resulted in a shorter postpartum interval to first ovulation but did not affect other reproductive measures in healthy, lactating dairy cows.     

Methionine availability in plasma of dairy cows supplemented with methionine hydroxy analog isopropyl ester

Adequate Met supply is especially important in the dairy cow for milk protein synthesis. Because of insufficient Met contents in the most frequently used feed-stuffs, Met becomes limiting in the diet of the dairy cow. To restore the amino acid balance of the diet and consequently to optimize lactation performance, Met must be supplied in a protected form because of its high degradability as a free amino acid by rumen microorganisms. A new chemical derivative of Met, the isopropyl ester of the 2-hydroxy-4-(methylthio)-butanoic acid (HMBi) was tested for its metabolic fate by following the evolution of plasma concentrations of its metabolites (2-hydroxy-4-(methylthio)-butanoic acid (HMB), Met, isopropyl alcohol, and acetone) after spot-dose supplementation (50 g Met equivalent) to 15 cows. Results indicated that HMBi would be quickly absorbed and hydrolyzed into HMB and isopropyl alcohol, and then converted to Met and acetone, respectively. In our experimental conditions, the Met availability for cows was estimated to be 48.34 ± 2.05% using a calibration curve established by modeling the area under the curve response to increasing doses of Met supplied as Smartamine M, whose bioavailability (80%) is considered the reference value. Plasma kinetics and bioavailability of Met were compared between HMBi and Smartamine M in the same cows. Comparison of the kinetics suggests that HMBi would be absorbed through the rumen wall providing good protection against rumen microorganisms. It can thus be concluded that HMBi is a new source of Met for ruminants with an acceptable bioavailability.     

Rumen degradation and availability of various amounts of liquid methionine hydroxy analog in lactating dairy cows

Ruminal escape of various amounts of methionine hydroxy analog [D,L-2-hydroxy-4-(methylthio)-butanoic acid (HMB)] was measured in an experiment designed as a 4 x 4 Latin square using four lactating dairy cows with cannula in the rumen and duodenum. The cows were fed a diet composed of corn silage, alfalfa haylage, rolled barley grain, canola meal, and blood meal, three times per day. The cows were fed the liquid analog each day for 1 wk before the experiment was started. On the day of the experiment, each cow received an intraruminal bolus dose of 0, 25, or 50 g of the liquid analog (Alimet feed supplement, 88% HMB) or 51.2 g of a dry calcium salt of the analog (86% HMB; MHA) mixed with 0.5 kg of ground barley grain. A liquid phase marker (Co-EDTA) was administered as a bolus dose into the rumen at the time of administration of the methionine hydroxy analogs. Rumen and duodenal contents, and blood serum were collected at 0, 1, 3, 6, 9, 12, and 24 h relative to the time of dosing. Rumen and duodenal samples were analyzed for Co and HMB, and serum was analyzed for free methionine. Fractional rate constants for the passage of the liquid marker (k(p)) and the decline of HMB concentration in the rumen (k(rHMB)) were determined by nonlinear regression. Liquid passage from the rumen was similar among the four analog treatments (0.136 +/- 0.012/h; mean +/- SEM). Ruminal escape of HMB as a percentage of the dose (100% x k(p)/k(rHMB)) did not differ between cows receiving 25, 50, and 51.2 g of the methionine analogs (42.5, 41.0, and 34.9 +/- 9.0%, respectively) and averaged 39.5%. Duodenal appearance of HMB as a percentage also did not differ between cows receiving 25, 50, and 51.2 g of the methionine analogs (16.2, 26.8, and 22.7%, respectively) and averaged 22%. Omasal absorption of HMB was variable ranging from 12.3 to 26.3% and averaged 17.6%. Serum methionine concentration peaked at 3 and 6 h after dosing and increased in proportion to the amount of the analog administered. It was concluded that 39.5% of the methionine hydroxy analog escaped rumen degradation, the percentage of the dose that escaped the rumen was not affected by the amount or form of the methionine analog fed, and the analog that escaped ruminal degradation was likely absorbed and metabolized to methionine.     

The effect of housing and methionine intake on hoof horn hemorrhages in primiparous lactating Holstein cows

This study investigated the effects of housing and nutrition on the development of hoof horn disease (as identified by the appearance of hoof horn hemorrhages) in primiparous lactating Holstein cows. The first objective was to investigate whether replacing butyl rubber mats in cubicles (free stalls) with thicker mattresses filled with chopped rubber would significantly reduce hoof horn hemorrhages, and if this reduction would so affect the level of hoof horn hemorrhages as to make it similar to that observed in primiparous cows in straw yards. The second objective was to investigate the effect of methionine supplementation for the first 13 wk of lactation on the development of such hemorrhages. This study confirmed that both sole and white line hemorrhages increase during early lactation in housed cows, although the pattern of development of white line hemorrhages is not identical to that of sole hemorrhages. Housing primiparous cows in straw yards after calving significantly reduced the development of hoof horn hemorrhages, but replacing cubicle mats with thicker mattresses had no significant effect. Providing 115% of calculated methionine requirements had no significant impact on the development of hoof horn hemorrhages.     

Supplementing limited methionine diets with rumen-protected methionine, betaine, and choline in early lactation Holstein cows

Eighty lactating Holstein cows from 21 to 91 d in milk were fed a corn silage-based total mixed ration (TMR) formulated with the Met content limited (42 g/ d) to investigate the impact of supplementing rumen-protected (RP) forms of Met, betaine, and choline on performance and metabolism. One of 4 supplements was blended into the TMR to produce 4 dietary treatments: 1) control, 2) 20 g/d of RP-Met, 3) 45 g/d of RP-betaine, and 4) 40 g/d of RP-choline. Calcium salts of fatty acids were used to protect both RP-betaine and RP-choline supplements. A similar amount of Ca salts of fatty acids was included in both control and RP-Met supplements to provide equal amounts of fat to all treatments. Overall, no differences in intake, milk yield, or milk composition were observed in primiparous cows. Average dry matter intake, body weight, and body condition score were not different among treatments in multiparous cows. Milk yield was higher in multiparous cows fed RP-choline compared with the other treatments. Multiparous cows fed RP-choline had higher milk protein yield than cows fed control or RP-betaine but was not different from cows fed RP-Met. Multiparous cows fed RP-choline had higher milk fat yield than cows fed RP-Met but was not different from cows fed control or RP-betaine. There were no beneficial effects of RP-betaine supplementation to a Met-limited TMR.     

Energy balance and lactation response in Holstein cows supplemented with cottonseed with or without calcium soap.

Holstein cows (n = 58, 21 primiparous), fed corn and wilted grass silages (63:37, DM basis) for free choice consumption, were assigned to control concentrate or supplemented concentrate during wk 1 to 16 postpartum with linted whole cottonseed (15% of projected DMI) alone or with Megalac (.54 kg/d). Our objective was to examine the effects of fatty acids on energy and N balances, total tract digestibility, and milk fatty acids in wk 7 and 16 and to assess total lactation responses. During balance measurements, fatty acids constituted 4.1, 6.8, and 8.6% of DM in control, oilseed, and oilseed plus protected fatty acid diets. Fat additions reduced fiber digestion (attributed to oilseed) and, to some degree, DMI and milk yield, but enhanced fat test without affecting protein percentage. Supplementary fat increased the proportion of C18:0 in milk at the expense of short-chain fatty acids. Supplemental oilseed with or without protected fatty acids reduced total heat production by 6% and reduced heat in excess of maintenance by 8%. Best estimates of NEL in linted whole cottonseed and of fat in Megalac were 1.81 and 5.69 Mcal/kg of DM. In total lactation, primiparous cows yielded more milk and FCM when fed oilseed plus Megalac and less of each when fed oilseed alone than controls. In pluriparous cows, milk yield was reduced by 2.7 kg/d relative to other treatments when oilseed plus Megalac was fed; FCM yield increased about 2 kg/d only when oilseed was supplemented alone. Overall, data suggest that basal ration fat and oilseed supplementation were too high or that supplementation should have been delayed until feed intake was higher.     

Metabolic responses of transition Holstein cows fed anionic salts and supplemented at calving with calcium and energy

The objective of this study was to determine the concentrations of plasma Ca, P, Mg, nonesterified fatty acids (NEFA), 3-hydroxybutyrate (BHBA), and glucose in transition cows fed anionic salts prepartum and provided with calcium and energy supplements at calving. The study was conducted on a Florida Holstein dairy farm from November to December 1997. Treatments consisted of no treatment (n = 30); 60 g of Ca as calcium chloride, orally (n = 30); 110 g of Ca as calcium propionate 510 g plus 400 g of propylene glycol, orally (n = 30); two doses of 60 g of Ca as calcium chloride, one at calving and the second 24 h later, orally (n = 30); and 10 g of Ca as borogluconate, intravenously (n = 30). Treatments were administered within 12 h after parturition. Blood samples were collected at d 1 (parturition), 2, 3, 6, 9, and 12 after calving. Plasma total Ca, P, Mg, NEFA, BHBA, and glucose were measured. There were no differences in the concentrations of the blood metabolites among treatments.     

Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 μg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO₂. Both betaine-homocysteine S-methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1β (IL1B), myeloperoxidase (MPO), IL10, and IL6] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS, GSR, IL1B, and IL6, suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response.     

Physiological responses to intramammary or intravenous treatment with endotoxin in lactating dairy cows

Twenty-one, middle to late lactation Holstein cows were assigned to one of three treatments in a completely randomized design to examine physiological changes associated with intramammary or intravenous administration of Escherichia coli endotoxin. Treatments were 1) Hank's balanced salt solution infusion in two contralateral quarters (control), 2) E. coli endotoxin infusion in two contralateral quarters, and 3) intravenous infusion of E. coli endotoxin. Blood was sampled and rectal temperature was measured at 30-min intervals. Endotoxin treatment was at 0900 h and sampling continued until 1700 h. Serum prolactin, cortisol, and 13,14-dihydro-15-keto-prostaglandin F2 alpha were measured. A pyretic response was observed in intravenous and intramammary treatment groups after endotoxin treatment. Response peak was higher (41.1 vs. 40.3 degrees C) and occurred later (6 vs. 4.5 h posttreatment) in the intramammary than the intravenous treatment group. Significant prolactin peaks were observed also in intravenous and intramammary endotoxin treatment groups. Prolactin peaked higher (288 vs. 112 ng/ml) and occurred sooner (1 vs. 4 h posttreatment) in the intravenous than in the intramammary treatment group. Cortisol followed a trend similar to prolactin. Cortisol peaked higher (100 vs. 82 ng/ml) and sooner (2.5 vs. 4.5 h posttreatment) in the intravenous than in the intramammary treatment group. Concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha increased rapidly posttreatment in the intravenous group only.     

Aluminosilicate clay improves production responses and reduces inflammation during an aflatoxin challenge in lactating Holstein cows

Mitigation strategies are vital in minimizing the health and economic risks associated with dairy cattle exposure to aflatoxin (AF). The objective of this study was to determine the effects of a commercially available aluminosilicate clay in a lactation diet on production responses, blood chemistry, and liver inflammatory markers of multiparous lactating Holstein cows during an AF challenge. Sixteen multiparous lactating Holstein cows [body weight (mean ± SD) = 758 ± 76 kg; days in milk = 157 ± 43 d] were assigned to 1 of 4 treatments in a replicated 4 × 4 Latin square design with 21-d periods: no adsorbent and no AF challenge (CON), no adsorbent and an AF challenge (POS), 113 g of aluminosilicate clay top-dressed on the total mixed ration (adsorbent; FloMatrix, PMI Nutritional Additives, Arden Hills, MN) with an AF challenge (F4), or 227 g of adsorbent with an AF challenge (F8). The challenge consisted of 100 μg of AFB₁/kg of dietary dry matter intake administered orally. For each period, milk was sampled 3× daily from d 14 to 21; blood, feces, and urine were sampled on d 14, 18, and 21; and liver samples were taken on d 18. Liver tissue was assessed for gene expression and histological hepatocyte inflammation. Statistical analysis was preformed using the MIXED and GLIMMIX procedures of SAS (SAS Institute Inc., Cary, NC). Fat-corrected milk (POS = 37.2, F4 = 39.2, and F8 = 38.9 kg/d) increased as concentration of adsorbent in the diet increased. There was a decrease in milk AFM₁ concentration at d 18 as concentration of adsorbent in the diet increased (POS = 0.33, F4 = 0.32, and F8 = 0.27 µg/kg). There was a decrease in AFM₁ concentration in urine (POS = 2.10, F4 = 1.89, and F8 = 1.78 µg/kg) and AFB₁ concentration in feces (POS = 4.68, F4 = 3.44, and F8 = 3.17 µg/kg) as concentration of adsorbent in the diet increased. Cows in CON had greater concentrations of serum cholesterol (202 mg/dL) and plasma superoxide dismutase (2.8 U/mL) compared with cows in POS (196 mg/dL and 2.6 U/mL, respectively). Plasma glutamate dehydrogenase increased as concentration of adsorbent in the diet increased (POS = 37.8, F4 = 39.3, and F8 = 39.1 U/L). The expression of NFKB1 was greater in the liver of cows in POS (0.78) compared with cows in CON (0.70). The expression of MTOR was greater in the liver of cows in CON (1.19) compared with cows in POS (0.96). When compared with cows in CON, cows in POS had greater odds ratio for hepatocyte inflammation (odds ratio = 5.14). In conclusion, the adsorbent used in this study had a positive effect on milk production and hepatocyte inflammation and reduced AF transfer.     

Effects of bovine somatotropin on physiologic responses of lactating Holstein and Jersey cows during hot, humid weather

Thirty-one lactating Holstein and Jersey cows were used to determine the effects of daily injections of 0 or 20 mg of recombinant bST on physiologic responses during hot, humid weather. Body temperature was determined by measuring milk temperature at each milking. Jugular blood was sampled for serum analysis of selected hormones, blood metabolites, and fatty acids, and arterial blood was sampled for blood pH and blood gas analysis. Milk was characterized for fatty acid composition. Blood pH was unchanged, but partial pressure of blood CO2, blood bicarbonate, base excess, and total CO2 declined with administration of bST. Serum triglycerides increased 89% in cows receiving bST. Blood urea nitrogen tended to decline in cows receiving bST. Serum cortisol, triiodothyronine, and thyroxine did not change, but insulin-like growth factor-1 increased 128% with bST use. Reduced milk short-chain fatty acids, increased milk long-chain fatty acids, and increased blood serum C18:1 fatty acid content occurred in cows administered bST and probably reflected tissue mobilization. Cows administered bST in hot weather had higher milk temperatures. Alterations in physiologic and metabolic measures in association with higher milk temperature suggest an interaction of bST use with hot, humid weather and reflect the need to minimize the effects of heat stress.     

Effects of rumen-protected methionine and other essential amino acid supplementation on milk and milk component yields in lactating Holstein cows

Objectives of this study were to investigate the effects of supplementing rumen-protected methionine (RP-Met), threonine (RP-Thr), isoleucine (RP-Ile), and leucine (RP-Leu) individually or jointly to a low-protein diet, on the performance of lactating dairy cows, as well as to determine the effects of these amino acids (AA) on the mammalian target of rapamycin (mTOR) in vivo. Ten lactating Holstein cows were randomly allocated to a repeated 5 × 5 Latin square experiment with five 19-d periods. Treatments were high-protein diet (16% crude protein, positive control; HP), low-protein diet (12% crude protein, negative control; LP), LP plus RP-Met (LPM), LP plus RP-Met and RP-Thr (LPMT), and LP plus RP-Met, RP-Thr, RP-Ile, and RP-Leu (LPMTIL). The dry matter intakes (DMI) of the LP, LPM, and LPMT diets were lower than that of the HP diet, whereas the DMI of the LPMTIL diet was intermediate between the HP diet and the other LP diets. Supplementing RP-Met to the LP diet increased the yields of milk and milk protein, increased the content of milk urea N, and tended to increase milk N efficiency. Co-supplementation of RP-Thr with RP-Met resulted in no further milk production increase. Co-supplementation of all 4 rumen-protected amino acids (RP-AA) increased milk and lactose yields to the level of the HP diet and tended to increase milk protein yield compared with the LPMT diet. We found no significant differences in the contents and yields of milk components between the LPMTIL and HP diets except for a lower milk urea N content in the LPMTIL diet. Venous concentrations of the measured AA were similar across the LP and LP diets supplemented with RP-AA. Relative to levels of the HP diet, LP diets had higher venous concentrations of Met and Gly and tended to have higher Phe concentration and lower concentrations of Val and BCAA. The LPMTIL diet had higher venous concentrations of Arg, Lys, Met, Phe, and Glu, and a lower Val concentration. Phosphorylation status of the measured mTOR components in LPM and LPMT treatments were similar to those in the LP treatment but phosphorylation status of mTOR and eIF4E-binding protein 1 (4eBP1) in LPMTIL treatment were higher. The phosphorylation rates of eukaryotic elongation factor 2 (eEF2) in the 4 LP and LP plus RP-AA diets were higher than that of the HP diet. Overall, results of the present study supported the concept that under the relatively short time of this experiment, supplementing RP-AA, which are believed to stimulate the mTOR signal pathway, can lead to increased milk protein yield. This increase appears to be due to increased DMI, greater mTOR signaling, and greater eEF2 activity.     

Wheat straw or alfalfa hay in diets with 30% neutral detergent fiber for lactating Holstein cows.

Twenty-four Holstein cows near peak lactation were fed diets formulated to contain 30% NDF with forage NDF from chopped wheat straw or chopped alfalfa hay in proportions of 0:3, 1:2, 2:1, and 3:0 in an 8-wk trial to evaluate effects of fiber source on lactational performance. Concentrate was based on highly degradable carbohydrate in steam-flaked sorghum grain. Forage to concentrate ratio decreased from 49:51 to 28:72 when straw was substituted for alfalfa hay. Milk yield (38.4 kg/d) and DMI (23.1 kg/d) were not influenced by forage source. Milk fat percentage decreased and milk protein percentage increased linearly with increasing straw. Yields of 3.5% FCM and fat showed linear and quadratic responses, because they appeared to be reduced only on the 0:3 diet, and protein yield showed a quadratic response and was highest for the 1:2 diet. Digestibility of NDF averaged 43.5, 45.4, 40.7, and 31.2%, and acetate to propionate ratio was 3.0, 2.4, 2.2, and 1.9 with increasing dietary straw. Passage rates were not influenced by diet and averaged 9.3, 7.6, 6.1, and 4.7%/h for fluid, grain, alfalfa hay, and wheat straw, respectively. It is suggested that the ratio of forage NDF to ruminally degradable starch be maintained greater than or equal to 1:1 when diets based on low quality forage are fed to cows in early lactation.     

Ruminal escape, gastrointestinal absorption, and response of serum methionine to supplementation of liquid methionine hydroxy analog in dairy cows

Availability of liquid methionine hydroxy analog [D,L-2-hydroxy-4-(methylthio)-butanoic acid] was evaluated in two experiments using four cannulated lactating dairy cows. The first experiment was designed as a 4 x 4 Latin square. Each cow received a pulse dose of 0, 30, 60, or 90 g of the methionine analog in the rumen each day for 10 d. Duodenal samples were collected at 16, 20, and 24 h after dosing for the last 5 d and pooled. The methionine analog was not detected in duodenal contents because it passed rapidly from the rumen relative to the sampling protocol. In the second experiment, cows were offered 90 g of the methionine analog and 600 ml of Cr-EDTA (3.5 g of Cr) mixed with ground corn for a period of 20 min after which any remains of the treatment were placed in the rumen. The concentration of the analog peaked in ruminal and duodenal fluid at 1 and 3 h, respectively. Based on the fractional rate constants for ruminal and duodenal disappearance of the methionine analog and passage of the liquid, it was determined that 50.0 +/- 2.8% of the methionine analog escaped ruminal degradation and became available for intestinal absorption (44.6 +/- 5.7%) or was absorbed from the omasum (5.4 +/- 3.3%). Serum methionine concentration peaked 6 h after analog dosing at a level that was three times the predose level, indicating that the methionine analog that escaped ruminal degradation was absorbed and metabolized to methionine.     

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.     

Production and nitrogen utilization in lactating dairy cows fed ground field peas with or without ruminally protected lysine and methionine

Previous research has shown that cows fed ≥24% of the diet dry matter (DM) as field peas decreased milk yield as well as concentration and yield of milk protein, possibly due to reduced DM intake and limited supply of Lys and Met. Twelve multiparous and 4 primiparous lactating Holstein cows were randomly assigned to 1 of 4 diets in a replicated 4 × 4 Latin square design. The diets contained (DM basis) 34.8% corn silage, 15.2% grass-legume silage, 5.9% roasted soybean, 2.4% mineral-vitamin premix, 2.0% alfalfa pellets, and either (1) 36% ground corn, 2.4% soybean meal, and 1.3% urea (UR), (2) 29.7% ground corn, 9.8% soybean meal, 0.13% ruminally protected (RP) Lys, and 0.07% RP-Met (CSBAA), (3) 25% ground field peas, 12.3% ground corn, and 2.4% soybean meal (FP), or (4) FP supplemented with 0.15% RP-Lys and 0.05% RP-Met (FPAA). Our objective was to test the effects of FP versus UR, FPAA versus CSBAA, and FPAA versus FP on milk yield and composition, N utilization, nutrient digestibility, ruminal fermentation profile, and plasma concentration of AA. Milk yield did not differ across diets. Compared with cows fed UR, those fed FP had greater DM intake, concentration and yield of milk true protein, apparent total-tract digestibility of fiber, urinary excretion of purine derivatives, and concentrations of total volatile fatty acids in the rumen and Lys in plasma, and less milk urea N and ruminal NH₃-N. The concentration of milk urea N, as well as the concentration and yield of milk fat increased in cows fed FPAA versus CSBAA. Moreover, cows fed FPAA had greater ruminal concentration of total volatile fatty acids, increased proportions of acetate and isobutyrate, and decreased proportions of propionate and valerate than those fed CSBAA. The plasma concentrations of His, Leu, and Phe decreased, whereas plasma Met increased and plasma Lys tended to increase in cows fed FPAA versus CSBAA. Concentration of milk true protein, but not yield, was increased in cows fed FPAA versus FP. However, cows fed FPAA showed decreased concentrations of His and Leu in plasma compared with those fed FP. Overall, compared with the CSBAA diet, feeding FPAA did not negatively affect milk yield and milk protein synthesis. Furthermore, RP-Lys and RP-Met supplementation of the FP diet did not improve milk yield or milk protein synthesis, but decreased urinary urea N excretion.     

Effect of forage source and a supplementary methionine hydroxy analog on nitrogen balance in lactating dairy cows offered a low crude protein diet

Four primiparous and 4 multiparous midlactation dairy cows were stratified by pre-experimental milk yield (23.5 ± 2.3 kg/d), protein yield (0.75 ± 0.066 kg/d), parity, and days in lactation (121 ± 10 d) into 4 groups of 2 in a 2 × 2 factorial, Latin square design (n = 8) to assess the effect of forage source and a supplementary methionine hydroxy analog on nitrogen (N) balance where low crude protein (CP) diets (13.3%) are offered. Diets contained either predominantly grass silage [GS (G− and G+)] or corn silage [CS (C− and C+)] as the forage source and were offered with (G+ and C+) or without (G− and C−) the isopropyl ester of 2-hydroxy-4 methylthio butanoic acid (HMBi). The G− and G+ contained 46% GS and 10% CS in the dry matter (DM), whereas C− and C+ contained 12% GS and 52% CS in the DM. Supplementary HMBi was included at a rate of 0.2% of DM in G+ and C+ diets. Diets were isonitrogenous (9.8 ± 0.4% protein truly digested in the small intestine) and isoenergetic (0.96 ± 0.01 units of energy for lactation; kg/DM). Each of the 4 experimental periods lasted 24 d: 14 d for dietary adaptation, followed by 10 d of housing in individual metabolism stalls; N balance was conducted on the last 5 d of each experimental period. Intake of DM was higher for CS-based vs. GS-based diets (20.23 vs. 18.41 kg/d). No effect of dietary treatment was found on milk yield or yields of milk fat, protein, and lactose. Supplementing with HMBi tended to improve milk solids yield (1.69 vs. 1.59 kg/d), casein yield (0.59 vs. 0.55 kg/d), and concentrations of casein (2.89 vs. 2.73%) and protein (3.58 vs. 3.49%) in the milk. Dietary N intake was higher for CS-based vs. GS-based diets (0.460 vs. 0.422 kg/d). However, forage source or supplementary HMBi had no effect on N excretion in the feces, urine, or milk. Excretion of urinary urea was positively related to N intake. Concentrations of urea N in the plasma (2.34 vs. 1.72 mmol/L), milk (2.54 vs. 2.24 mmol/L), and urine (123.32 vs. 88.79 mmol/L), and total excretion of urinary urea N (40.23 vs. 35.09 g/d) were higher for animals offered CS-based vs. GS-based diets. Corn silage improved N intake through improved DM intake. However, neither forage source nor HMBi supplementation affects N output in the feces, urine, or milk.     

Assessing amino acid uptake and metabolism in mammary glands of lactating dairy cows intravenously infused with methionine,lysine, and histidine or with leucine and isoleucine

The objective of this study was to evaluate the effect of jugular infusions of 2 groups of AA on essential AA (EAA) transport and metabolism by mammary glands. Four Holstein cows in second lactation (66 ± 10 d in milk) were used in 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Treatments were jugular infusions of saline; Met, Lys, and His (MKH); Ile and Leu (IL); or MKH plus IL (MKH+IL). Each period consisted of 8 d of no infusion followed by 8 d of jugular vein infusion of the treatment solutions. Amino acids were infused at rates of 21 g of Met, 38 g of Lys, 20 g of His, 50 g of Leu, and 22 g of Ile per day. Cows were fed a basal diet consisting of 15.2% crude protein with adequate rumen degradable protein but 15% deficient in MP based on estimates by Cornell Net Carbohydrate and Protein System (v6.5). On the last day of each period, ¹³C-AA derived from algae was infused into the jugular vein over 6 h, and blood and milk samples were collected before, during, and after infusion. Plasma and milk samples were analyzed for AA isotopic enrichment, and a mammary compartmental model was fitted to the data to derive bidirectional transport and metabolism rates for individual EAA. Influx of Leu increased with IL, whereas influx of other EAA was not different among treatments. Cellular efflux of Met and Lys to venous plasma represented 12 to 34% of influx, whereas cellular efflux of Phe and BCAA represented 29 to 59% of influx. Increased efflux/influx ratios of Ile and Leu with IL but not Met and Lys with MKH demonstrated that increased Ile and Leu influx was mostly returned to plasma resulting in no change in net uptake or efficiency. The isotope results showed that mammary net uptake of Lys and Ile increased during MKH infusion. Net uptake of Met increased with MKH but only in the absence of IL. Catabolism of Lys and Met only increased with MKH alone, resulting in decreased efficiency for milk protein, which demonstrated that Ile and Leu infusion can spare Lys and Met for milk protein synthesis. Total AA uptake to milk output was not different from 1, implying the catabolized Met and Lys contributed nitrogen to nonessential AA. Overall, EAA uptake and metabolism in mammary glands of dairy cows varied across individual EAA and responded differently to respective AA supplements. In addition, uptake, retention, and end use of AA by mammary tissue is variable and dependent on the mix of AA provided. This variability, depending on the mix of AA absorbed, will change the efficiency of utilization of individual AA at the mammary gland level and consequently the whole-body level. Thus, it is inaccurate to use a fixed, constant efficiency within and across AA to represent tissue activity.     

Production,physiological response,and calcium and magnesium balance of lactating Holstein cows fed different sources of supplemental magnesium with or without ruminal buffer

The objective of this study was to evaluate the effects of dietary replacement of magnesium oxide (MgO) with calcium-magnesium hydroxide [CaMg(OH)₂] and its interaction with ruminal buffer (sodium sesquicarbonate) supplementation on production, Ca and Mg balance, and overall physiological response of mid-lactation Holstein dairy cows. Sixty cows averaging 40.5 ± 7.0 kg of milk/d were used. Treatments were assigned following a 2 × 2 factorial arrangement: (1) MgO, (2) MgO + buffer, (3) CaMg(OH)₂, or (4) CaMg(OH)₂ + buffer. Diets were formulated to have 16.5% of crude protein, 1.82 Mcal/kg of net energy for lactation, 0.67% Ca, 0.39% P, and 0.25% Mg, all on a dry matter (DM) basis. Treatments were individually top dressed. Milk production, composition, and DM intake were evaluated. A subsample of 20 cows were randomly selected for the evaluation of Ca and Mg balance, blood gases, and electrolytes. Ruminal fluid was also collected for evaluation of pH and Ca and Mg solubility. Effects of Mg source, buffer, and the interaction Mg source × buffer were analyzed through orthogonal contrasts. An interaction of Mg source × buffer was found for DM intake and feed efficiency, in which cows fed CaMg(OH)₂ had a similar feed efficiency regardless of ruminal buffer inclusion; however, when cows were fed MgO, the inclusion of buffer reduced feed efficiency. No effects on body weight and milk yield were observed. Buffer addition tended to increase the concentrations of fat, protein, and solids-not-fat, without affecting the yields of these milk components. Magnesium source and buffer did not affect ruminal fluid, blood, urine, or fecal pH; however, buffer supplementation increased urinary pH. Treatment with CaMg(OH)₂ increased blood concentration of HCO₃^?, total CO₂, and base excess compared with cows fed MgO. No differences were observed in the ruminal solubility of Ca and Mg or on milk or urinary Ca and Mg excretion. Greater plasma Mg concentration was observed for animals fed MgO compared with cows fed CaMg(OH)₂; however, both sources were above the threshold recommended in the literature for dairy cows. Also, a reduction in fecal Mg excretion was observed in animals fed CaMg(OH)₂. In summary, we provide evidence that CaMg(OH)₂ could replace MgO without affecting performance, overall physiological response, or Ca and Mg balance of mid-lactating dairy Holstein cows.     

火焰原子吸收光谱法测定铜锤草中锰铁铜锌

铜锤草样品经浓硝酸-高氯酸(3+1)混合酸消解,用火焰原子吸收光谱法测定了其不同部位锰、铁、铜和锌四种金属元素的含量。测定工作曲线相关系数R在0.9958-0.9993之间,加标回收率在94.80%-107.91%之间,相对标准偏差均小于1.78。测定方法简单易行,结果可靠。