Effects of supplementation with yeast culture and enzymatically hydrolyzed yeast on performance of early lactation dairy cattle.

One hundred fifty multiparous cows were balanced to 1 of 3 treatments (2 pens/trt) according to previous lactation 305-d mature equivalent yield to evaluate supplementation with yeast culture (YC; A-Max, Vi-COR, Mason, IA) and YC plus enzymatically hydrolyzed yeast (YC+EHY; Celmanax, Vi-COR) on production performance in dairy cattle. Cows entered pens at calving and remained through 14 wk postpartum. Treatment assignment to pens was random throughout the barn. Pens were identical in layout and each contained an exit alley to eliminate feed and animal mixing. The 3 treatments were control: nonsupplemented; YC: control diet with YC (56 g/d); and YC+EHY: control diet plus YC and EHY (28 g/d). Mean pen dry matter intake was similar across treatments. Cows supplemented with YC and YC+EHY produced more milk, fat-corrected milk, and energy-corrected milk than control cows (1.4 and 1.6, 1.6 and 1.8, 1.7 and 1.9 kg, respectively). Treatments YC and YC+EHY did not differ. Milk fat and lactose percentages were not affected by treatment. Milk protein percentage was higher for cows supplemented with YC+EHY than for those on YC and control treatments (2.98, 2.93, and 2.91%, respectively) with control and YC-supplemented cows not being different from each other. Differences in fat and protein yields were primarily reflective of milk yield. Treatment had no effect on milk urea nitrogen. No differences in the incidence of metabolic health were observed; however, cases of clinical mastitis for YC+EHY were less than half those for control and YC during wk 8 to 14 on trial. Somatic cell count was higher for cows fed control and YC diets compared with YC+EHY, primarily during wk 8 to 14 on trial. Supplementation of early lactation cows with YC improved milk production performance; furthermore, EHY supplementation improved milk protein percentage and mammary gland health.

Effects of feeding a dietary antioxidant in diets with oxidized fat on lactation performance and antioxidant status of the cow.

The objective of the study was to evaluate the effect of feeding the dietary antioxidant Agrado Plus (AOX; Novus International, St. Louis, MO) in diets that contained 2% fresh fat (FF) or oxidized fat (OF) on milk production and composition and antioxidant status of cows during mid to late lactation. Forty-four mid to late lactating primiparous cows housed in a tie-stall barn were fed a diet that contained 2% FF for 15 d as adaptation period and then randomly allocated to 1 of the 4 dietary treatments (FF, FF+AOX, OF, OF +AOX) for 6 wk. Feeding AOX increased dry matter intake, 3.5% fat-corrected milk, and milk fat yield, and decreased milk protein content but not yield. Feeding OF increased milk fat yield, but decreased dry matter intake and body weight gain. Milk fat composition changed with treatments: AOX increased cis 18:1 and decreased trans-11 18:1, whereas OF decreased trans-9 and trans-11 18:1 and cis-9, trans-11 18:2 in milk. Plasma antioxidant enzymes and status were affected by treatments. Feeding OF increased superoxidase dismutase activity but decreased plasma antioxidant status, whereas AOX supplementation increased glutathione peroxidase activity across fat types and increased the antioxidant status and superoxidase dismutase activity when feeding OF diets. It can be concluded that feeding AOX improved lactation performance and the antioxidant status of the cow across fat types, and feeding OF increased milk fat yield but decreased dry matter intake, body weight gain, and antioxidant status. The negative effects of feeding OF were partially alleviated by AOX.

Direct-fed microbial supplementation and health and performance of pre- and postpartum dairy cattle: a field trial.

A double-blind field trial was conducted on a commercial dairy to study the effects of feeding a direct-fed microbial (DFM) product consisting of 2 strains of Enterococcus faecium plus Saccharomyces cerevisiae yeast on prepartum and postpartum performance of Holstein cows. Treatments consisted of the normal pre- and post-fresh TMR supplemented with the DFM (2 g/cow per d) or a placebo. Treatments started approximately 10 d prepartum and continued until about 23 d in milk (DIM). A total of 366 Holstein cows were enrolled in 1 of 2 placebo groups or 2 DFM-supplemented groups. Groups were enrolled consecutively, starting with the placebo treatment. Sample size was limited to 4 groups because the cooperating dairy prematurely terminated the study due to increased health problems in one of the groups. Blood samples were taken during the prefresh period between 2 and 10 d prior to calving and at weekly intervals from 3 to 23 DIM. Blood concentrations of nonesterified fatty acids before calving and beta-hydroxy-butyrate after calving were not affected by treatment. Supplementation with the DFM product increased milk fat percentage for the first lactation cows and increased milk protein percentage for the second and greater lactation cows during the first 85 DIM. Second-lactation cows fed the DFM product received fewer antibiotic treatments before 85 DIM than cows receiving the placebo. This validated the dairy producer's concern that cows consuming one of the diets (revealed to be the placebo diet after the study was completed) were experiencing more health problems. Most measures of milk yield were numerically increased by supplementation with the DFM product. However, differences in milk yield were not significant. Key covariates for main study outcomes included milk yield in the previous (first) lactation, body condition score prior to calving, days spent in the maternity pen, and stocking density of the pre-fresh pen.

The effect of trace mineral fortification level and source on performance of dairy cattle.

Five hundred seventy-three cows, balanced by parity and 305-d mature equivalent at dry off, were assigned to 1 of 4 treatments: 1) 75% complexed trace minerals (CTM; 75C): Zn, Mn, Cu, and Co supplied at 75% of NRC (2001) guidelines by Zn-, Mn-, and Cu-specific AA complexes, and cobalt glucoheptonate; 2) 100% inorganic (100I): Zn, Mn, Cu, and Co supplied at 100% of NRC (2001) requirements by sulfate sources; 3) 100% complexed (100C): Zn, Mn, Cu, and Co supplied at 100% of NRC (2001) requirements by CTM; and 4) complexed/ inorganic (C/I): Zn and Cu supplied at 100% of NRC (2001) requirements using a combination of CTM and sulfates and Co and Mn supplied with sources at 9.1 and 3.3 times NRC (2001) requirements using a combination of CTM and sulfates. All percentages of Zn, Cu, Mn, and Co relative to NRC (2001) reflect supplemental contributions and do not include basal diet contributions. Experimental periods were dry period 1, full lactation 1, dry period 2, and 200 d into the subsequent lactation. Reproductive, health, and production information was collected during both lactations. Claw evaluations were conducted at trial start, 150 d into lactation 1, at the end of lactation 1, and 150 d into lactation 2. During lactation 1, C/I cows produced more milk, fat-corrected milk, energy-corrected milk, and fat than 100I cows. During lactation 2, yields of milk, fat-corrected milk, energy-corrected milk, fat, and protein were higher for 100C and C/I cows than for 75C or 100I cows. Fat percentage was highest for 100C cows with no treatment effect on protein content. During lactations 1 and 2, C/I cows had fewer days to first estrus than cows receiving the other treatments. During lactation 2, C/ I cows had fewer services per conception and days open. There were no significant effects of treatment on health. White line separation incidence was lower for 100I cows than 75C cows, whereas heel erosion was higher for the 100I cows than for the C/I cows. Fortification of trace elements with inorganic and complexed sources at or above NRC requirements improved reproductive and productive performance. In addition, cows can be supplemented with CTM at 75% of NRC requirements with no reduction in performance compared with supplementing at 100% of NRC requirements using only sulfate sources of Zn, Mn, Cu, and Co.

Direct-fed microbial supplementation on ruminal digestion, health, and performance of pre- and postpartum dairy cattle.

Effects of supplementing direct-fed microbial agents (DFM) to dairy cows during the transition period were evaluated. Forty-four Holstein cows were fed close-up and lactating diets that did or did not contain 2 g of DFM/cow per d. The direct-fed microbial (DFM) supplement was fed at a rate of 2 × 109 viable yeast cells and 5 × 109 cfu of bacteria per cow per day [corrected].Supplemented cows were fed the DFM 21 d prior to expected calving date through 10 wk postpartum. Cows supplemented with DFM had higher estimated ruminally available dry matter (DM) for both corn silage and haylage than did control cows. Supplemented cows consumed more DM during both the pre- and postpartum periods. In addition, those supplemented with DFM produced 2.3 kg more milk/cow per d than did nonsupplemented cows. There was no difference in 3.5% fat-corrected milk. Milk fat percentage was lower, but not depressed (4.76 vs. 4.44%) for cows receiving DFM. There were no differences in milk fat yield or milk protein percentage and yield. Cows consuming DFM had higher blood glucose postpartum, as well as lower beta-hydroxybutyrate levels both prepartum and on d 1 postpartum. Plasma nonesterified fatty acid concentration was not statistically affected by DFM, but was numerically lower prepartum and higher postpartum for supplemented cows. This study demonstrated that targeted DFM supplementation enhanced ruminal digestion of forage DM. Early lactation cows receiving supplemental DFM produced more milk and consumed more DM during the pre- and postpartum periods. Cows consuming DFM, however, experienced a lower, but not depressed, fat percentage compared with nonsupplemented cows.

Direct-fed microbial supplementation on the performance of dairy cattle during the transition period.

The influence of a direct-fed microbial (DFM) on the prepartum period and the effects on production performance during the postpartum period was investigated using 64 multiparous Holstein cows. Two close-up dry cow diets were fed to two groups of 32 cows each starting 21 d precalving as follows: 1) no DFM and 2) DFM. Post-calving cows were fed a lactation ration with or without DFM supplementation.The direct-fed microbial (DFM) supplement contained 2 × 109 viable yeast cells and 5 × 109 cfu of bacteria (Enterococcus faecium) per cow per day, top dressed in a 90-g supplement [corrected].The DMI during the prepartum period was not affected by DFM supplementation. During the postpartum period, DMI, milk yield, and milk protein content were higher for cows receiving DFM supplementation compared with no DFM. Blood glucose and insulin levels were higher and NEFA levels were lower for cows receiving DFM during the postpartum period. These data suggest that targeted DFM supplementation increased DMI and milk production postpartum. Blood metabolite information would suggest this response was associated with more glucose being made available and less fatty acids being mobilized from lipid stores.

Influence of yeast culture on ruminal microbial metabolism in continuous culture.

A continuous culture study was conducted to evaluate the effect of two different yeast cultures on ruminal microbial metabolism. The treatments were a) control lactation ration, b) yeast culture 1 (YC1, Diamond-V XP) and c) yeast culture 2 (YC2, A-Max), both fed at an equivalent of 57 g/head per day. The results showed that both yeast culture products increased dry matter (DM) digestion, propionic acid production, and protein digestion compared with the control. Yeast culture 1 demonstrated an increase in molar percentage of propionic acid, a reduction in acetic acid, and a lower mean nadir (daily low) pH compared with YC2. Ruminal cultures treated with YC digested more protein and contributed less bypass N than control. Supplementing YC2 resulted in a tendency for higher microbial N/kg DM digestion than YC1. Yeast culture 1 resulted in production of rumen microbes containing less protein and more ash than YC2. These results support previous research findings that yeast culture does influence microbial metabolism, and specific yeast cultures may have different modes of action.

Evaluation of an indwelling ruminal probe methodology and effect of grain level on diurnal pH variation in dairy cattle.

To evaluate a ruminal probe for recording diurnal variation in rumen pH, we fitted three ruminally fistulated cows with probes affixed to the inside of the cannula. Probes were connected to a data logger and readings were recorded hourly. The experiment was a Latin square design with cows fed three different diets: 50, 60, and 70% grain as a total mixed rations once daily. Feed offerings and refusals were recorded daily. The experimental period was 21 d. The first 6 d were for adaptation, followed by 5-d rotations through each of the diets. Daily probe readings were recorded at 4-, 6-, 8-, 12-, and 24-h intervals. At each interval, readings were recorded (precleaned), a sample of rumen fluid was taken, and pH was measured in the laboratory. As probes were removed from the rumen, probe ends were cleaned with 0.1 N HCl and reinserted into the rumen, and a reading was recorded (postcleaned). No protective pH probe shields were used in this experiment. There were no differences between pre- and postcleaned pH readings across cows for all diets. Mean time under pH 5.0, 5.5, and 6.0 were 0.2, 2, and 7.2 h, respectively. Diet affected length of time under a certain pH, only for readings under pH 6. Diurnal pH profiles were monophasic in nature. The degree of acidity increased after feeding and duration of nadir increased with increasing grain in the diet. Daily DMI increased but was highly variable within the first week after switching to the next higher grain level. These results indicate the use of an indwelling ruminal probe without a protective shield, cleaned, and calibrated daily can accurately measure diurnal variation of ruminal pH. In addition, transition to higher grain levels in the diet increases pH, duration of pH nadir, and daily DMI fluctuation.

Ruminal supplementation of direct-fed microbials on diurnal pH variation and in situ digestion in dairy cattle.

To evaluate the effect of direct-fed microbial (DFM) concentration on diurnal rumen pH profiles and in situ digestibilities, nine ruminally cannulated cows in early lactation were fed treatments consisting of DFM (Enterococcus faecium, Lactobacillus plantarum, and Saccharomyces cerevisiae) at a level of a) 1 x 10(5) cfu/ml of rumen fluid (10(5)), b) 1 x 10(6) cfu/ml of rumen fluid (10(6)), and c) 1 x 10(7) cfu/ml rumen fluid (10(7)). Treatments were directly administered via rumen cannula once daily. Cows were fitted with pH probes in their cannula and connected to dataloggers, which monitor pH hourly. The experimental period was 21 d: 7-d adjustment, 14-d for pH, and in situ measurements. Cows fed 10(5) were able to sustain a higher nadir pH than were cows fed 10(6) or 10(7). Cows fed 10(5) had a higher digestion rate of high moisture ear corn (HMEC) dry matter. Corn silage digestion was higher for cows fed 10(5) and 10(6) compared with those receiving 10(7). There were no carryover effects of treatment associated with rumen pH when switching from one treatment regimen to the next. Results from this study demonstrate that incorporation of a specific level of DFM aids in reducing diurnal ruminal acidity.

Digital characteristics in commercial dairy herds fed metal-specific amino acid complexes.

Five commercial dairy herds in Central New York fed metal-specific amino acid complexes were selected to evaluate digital characteristics. During the first year (period 1), herds were evaluated by a specific procedure by one individual. Three herds were not supplemented and two herds were supplemented with zinc methionine. During the subsequent year (period 2) all herds were switched to a combination of zinc methionine, copper lysine, manganese methionine, and cobalt glucoheptonate and evaluated in the same way as during period 1. There was no effect of period on incidence of heel erosion and interdigital dermatitis. During period 2, there was a reduced incidence of double soling, white line separation, sole hemorrhages, sole ulcers, and papillomatous digital dermatitis, and the incidence of wall ridges tended to be reduced compared with period 1. There was no effect of period on the incidence of abaxial wall lesions, digital arthritis, or foot rot, although the overall incidence of these disorders was low. During period 2, when cows were fed a combination of complexed trace minerals, there was a general reduction in the incidence of digital disorders associated with the laminitis syndrome complex compared with period 1; however, other time-related differences existed that may confound interpretation.

Extension of transverse relaxation-optimized spectroscopy techniques to allosteric proteins: CO- and paramagnetic fluoromet-hemoglobin [beta (15N-valine)].

We present the first steps in applying transverse relaxation-optimized spectroscopy (TROSY) techniques to the study of allosterism. Each beta-chain of the hemoglobin (Hb) tetramer has 17 valine residues. We have (15)N-labeled the beta-chain Val residues and detected 16 of the 17 (1)H-(15)N correlation peaks for beta-chain Val of the R state CO-Hb structure by using the TROSY technique. Sequence-specific assignments are suggested, based mainly on analysis of the (1)H pseudocontact-shift increments produced by oxidizing the diamagnetic R state HbCO to the paramagnetic R state fluoromet form. When possible, we support these assignments with sequential nuclear Overhauser effect (NOE) information obtained from a two-dimensional [(1)H,(1)H]-NOESY-TROSY experiment (NOESY, NOE spectroscopy). We have induced further the R-T conformational change by adding the allosteric effector, inositol hexaphosphate, to the fluoromet-Hb sample. This change induces substantial increments in the (1)H and (15)N chemical shifts, and we discuss the implication of these findings in the context of the tentative sequence assignments. These preliminary results suggest that amide nitrogen and amide proton chemical shifts in a selectively labeled sample are site-specific probes for monitoring the allosteric response of the ensemble-averaged solution structure of Hb. More important, the chemical-shift dispersion obtained is adequate to permit a complete assignment of the backbone (15)N/(13)C resonances upon nonselective labeling.

Bovine acidosis: implications on laminitis.

Bovine lactic acidosis syndrome is associated with large increases of lactic acid in the rumen, which result from diets that are high in ruminally available carbohydrates, or forage that is low in effective fiber, or both. The syndrome involves two separate anatomical areas, the gastrointestinal tract and body fluids, and is related to the rate and extent of lactic acid production, utilization, and absorption. Clinical manifestations range from loss of appetite to death. Lactic acid accumulates in the rumen when the bacteria that synthesize lactic acid outnumber those that utilize lactic acid. The systemic impact of acidosis may have several physiological implications, including laminitis, a diffuse aseptic inflammation of the laminae (corium). Although a nutritional basis for the disease exists, etiology includes a multitude of interactive factors, such as metabolic and digestive disorders, postpartum stress, and localized trauma, which lead to the release of vasoactive substances that trigger mechanisms that cause degenerative changes in the foot. The severity of laminitis is related to the frequency, intensity, and duration of systemic acidotic insults on the mechanisms responsible for the release of vasoactive substance. The critical link between acidosis and laminitis appears to be associated with a persistent hypoperfusion, which results in ischemia in the digit. Management of acidosis is critical in preventing laminitis. High producing dairy herds attempting to maximize energy intake are continually confronted with subclinical acidosis and laminitis. Management of feeding and husbandry practices can be implemented to reduce incidence of disease.

Inhibitor-enhanced electron transfer: copper cytochrome c as a redox-inert probe of ternary complexes.

Copper-substituted cytochrome c (CuCc) has been used as a structurally faithful, redoxinert inhibitor to probe the mechanism of electron transfer (ET) between Cc molecules and cytochrome c peroxidase (CcP). This inhibitor enhances photoinduced ET quenching of the triplet excited state of a zinc-substituted protein (ZnCcP or ZnCc) by its iron(III) partner (Fe3+Cc or Fe3+CcP). These results show that CcP and Cc form a ternary complex in which one Cc molecule binds tightly at a surface domain of CcP having low ET reactivity, whereas the second Cc molecule binds weakly to the 1:1 complex at a second domain with markedly greater (approximately 10(3)) reactivity. These results also rule out the possibility that Cc bound at the second domain cooperatively enhances ET to Cc at the first domain. The multiphasic kinetics observed for the photoproduced ET intermediate do not reflect electron self-exchange between two Cc molecules within the ternary complex.

Effects of a colostrum substitute and of dietary brewer's yeast on the health and performance of dairy calves.

We tested the effects of substituting a whey protein concentrate for dam's colostrum at first feeding and of 1% brewer's yeast in dry feed on calf health and performance. Eighty-four calves were assigned to four preweaning and eight postweaning treatments based on feeding either dam's colostrum or a whey protein concentrate at birth and either yeast or no yeast in the dry feed both before and after weaning. After the first feeding, all calves were fed colostrum through 4 d of age and an all milk protein milk replacer from d 5 to 46 of age. Daily feed intakes, health measurements, and weekly BW were recorded. Calves fed whey protein concentrate at birth had similar health parameters and greater feed efficiency than calves fed colostrum over the preweaning period. Addition of 1% brewer's yeast to the dry feed apparently reduced the incidence of fever and the number of associated antibiotic treatments during the preweaning period, but had no effects thereafter. Under the conditions of this study, whey protein concentrate was an effective substitute for dam's colostrum at first feeding, and 1% brewer's yeast fed in dry feed may reduce susceptibility to infection prior to weaning.

Feeding sequence and strategy effects on ruminal environment and production performance in first lactation cows.

Three primiparous cows were assigned randomly to a 4 x 4 incomplete Latin square to evaluate ruminal characteristics (Experiment 1) with the following feeding strategies: strategy A, supplement (protein), forage plus high moisture ear corn (energy), protein; strategy B, energy plus protein, forage, energy plus protein; strategy C, forage plus energy, protein, forage plus energy, protein; and strategy D, forage, energy plus protein, forage, energy plus protein. In Experiment 2, 36 cows were fed diets similar to those in Experiment 1 from wk 4 through 19 postpartum. Cows fed diets according to strategy A had the highest and strategy D the lowest mean ruminal pH. Diurnal variation in ruminal pH was less with cows fed diets using strategies A and B than C and D. Other ruminal parameters were not significantly different. In Experiment 2, cows fed according to strategy B had higher DMI than those fed according to other strategies. Energy and protein fed together and separately from forage, regardless of sequence, increased milk fat percentage compared with protein fed separately. Milk production and protein yield tended to be highest when protein was fed separately, regardless of sequence to forage. Strategic feeding of protein and energy sources in relation to forage influenced some ruminal and production measurements in primiparous cows.

Site of digestion of starch in the gastrointestinal tract of dairy cows and its effect on milk yield and composition.

Physical and chemical processing of feed ingredients and feeding management strategies are major instruments of manipulating amount and site of starch digestion in the gastrointestinal tract. Generally, as rumen escape of starch increases, postruminal starch digestion increases, and there does not appear to be a limitation to intestinal starch digestion. However, the efficiency with which postruminal starch is digested decreases, which represents a limitation that warrants investigation. Even though digestible dietary starch is presented to the intestine, there is no net glucose absorption at the portal vein, and plasma glucose levels remain relatively unaffected. This result may be associated with the large metabolic requirement for postruminally absorbed glucose, which is preferentially used for oxidative metabolism at the visceral tissue level. In addition, peripheral glucose concentration is highly regulated. A possible implication is that the exogenous glucose supply may spare endogenously synthesized glucose for gut metabolism, allowing more to be directed to the mammary gland. Amino acids also may be spared (less metabolism of dietary and tissue amino acids in the gut). Current production studies yield no clear evidence as to the benefits of postruminal digestion of starch to enhance milk yield or to change its composition. However, studies suggest that starch digested postruminally is used more efficiently for milk synthesis than that digested in the rumen.

Structural and functional significance of inhomogeneous line broadening of band III in hemoglobin and Fe-Mn hybrid hemoglobins.

Near-infrared spectra of hemoglobin and Fe-Mn hybrid hemoglobins have been obtained at cryogenic temperatures. The charge-transfer (a2u(pi)----dzy) transition at approximately 760 nm (band III) has been found to be a conformationally sensitive indicator of the heme-pocket geometry in these species. Temperature, protein tertiary and quaternary structure, chain heterogeneity, and ligand rebinding subsequent to CO photolysis all affect the line width and position of this transition. We conclude that the overall line shape of band III is derived from both subunit heterogeneity and conformational disorder within each subunit. A model is suggested that relates the observed pH dependence of the kinetic hole burning due to ligand rebinding to specific structural parameters of the proximal heme pocket that influence both the peak position and the inhomogeneous line shape of band III.

Nitric oxide adducts of the binuclear iron site of hemerythrin: spectroscopy and reactivity.

Nitric oxide forms adducts with the binuclear iron site of hemerythrin (Hr) at [Fe(II),Fe(II)]deoxy and [Fe(II),Fe(III)]semimet oxidation levels. With deoxyHr our results establish that (i) NO binds reversibly, forming a complex which we label deoxyHrNO, (ii) NO forms a similar but distinct complex in the presence of fluoride, which we label deoxyHrFNO, (iii) NO is directly coordinated to one iron atom of the binuclear pair in these adducts, most likely in a bent end-on fashion, and (iv) the iron atoms in the binuclear sites of both deoxyHrNO and deoxyHrFNO are antiferromagnetically coupled, thereby generating unique electron paramagnetic resonance (EPR) detectable species. The novel EPR signal of deoxyHrNO (deoxyHrFNO) with g[[ = 2.77 (2.58) and g = 1.84 (1.80) is explained by the magnetic interaction of the Fe(II) (S' = 2) and [FeNO]7 (S = 3/2) centers observed by Mössbauer spectroscopy. Antiferromagnetic coupling leads to a ground state of Seff = 1/2. Analysis of the EPR parameters using the isotropic spin-exchange Hamiltonian, Hex = 2JS3/2.S2, and including zero-field splitting leads to a coupling constant, -J approximately 23 cm-1, for deoxyHrNO. The resonance Raman spectrum of deoxyHrNO shows features at 433 and 421 cm-1 that shift downward with 15N16O and that are assigned to stretching and bending modes, respectively, of the [FeNO]7 unit. Sensitivity of the bending mode to D2O suggests that bound NO participates in hydrogen bonding. We propose that the terminal oxygen atom of NO is hydrogen bonded to the proton of the mu-hydroxo bridge in the Fe-(OH)-Fe unit. A bent Fe-N-O geometry is supported by spectroscopic and structural comparisons to synthetic complexes and is consistent with a limiting [FeII,FeIIINO-] formulation for deoxyHrNO. Reversibility of NO binding to deoxyHr is demonstrated by bleaching of the optical and EPR spectra of deoxyHrNO upon additions of excess N3- or CNO-. DeoxyHrNO undergoes autoxidation under anaerobic conditions over the course of several hours. The product of this autoxidation appears to be an EPR-silent NO adduct of semimetHr. The formal one-electron oxidations of the binuclear iron site of deoxyHr by NO and by HNO2 can conceivably occur with no net change in charge on the iron site. In contrast, autoxidation of oxy- to metHr requires a change in net charge on the iron site, which may provide a kinetic barrier.

Characterization of in situ dry matter and nitrogen digestion of various corn grain forms.

Corn was harvested and stored as either dried shelled, dried ear, high moisture shelled, or high moisture ear. Shelled corns were rolled and ear corns were ground through a forage harvester. High moisture corns were stored in sealed drums. Each corn type was subjected to in situ ruminal digestion with incubation times through 100 h for both original forms and ground at 5 mm. Dry matter, N, and nonprotein DM digestion rates were determined. Unground dried shelled corn possessed the fastest and high moisture ear corn the slowest rate of DM digestion; however, extent of digestion was greatest for high moisture corns within 16 h of ruminal exposure. Grinding increased the rate of shelled corn digestion. Nitrogen digestion rate was fastest for high moisture ear corn and slowest for dried shelled corn. Grinding increased the rate of nitrogen digestion for all corn forms except high moisture shelled corn. Generally, correction for bacterial nitrogen contamination resulted in increased rate of nitrogen digestion. Dried shelled corn possessed the fastest rate of nonprotein DM digestion. Ratio of nitrogen to nonprotein DM rates ranged from .56 to 1.24 for dried shelled corn and high moisture ear corn, respectively. Grinding narrowed this range. Digestion rates of corn forms may not necessarily reflect initial (1 to 16 h) extent of ruminal nutrient availability.

Description and validation of an in situ autoperfusion method to determine nutrient absorption and metabolism in bovine small intestine.

Holstein bull calves, 8 to 12 wk of age, were anesthetized with halothane gas. An approximate 20-cm section of small intestine, 60 to 90 cm proximal to the ileocecal junction was clamped to isolate blood circulation to a single set of arcuate vessels and to form an intestinal segment fitted for infusion and drainage. The vein was catheterized to allow total venous collection. Donor blood was transfused via jugular vein to replace venous drainage. This technique was evaluated in four calves by exposing the lumen to eight replications (12 or 20 min incubation, 30-min wash with 39 C saline) of 16 mM L-Met (14C-labeled). Time course appearance of Met in venous blood indicated similar rates and patterns of absorption for individual calves. There were no clinically significant alterations in jugular blood chemistry profiles across replications. Four calves were used to evaluate the effect of three isotonic perfusion media (saline, Krebs-Ringer bicarbonate and M-199 tissue culture media) on Lys and Met absorption. Venous flow rates and absorption of Lys were faster with Krebs buffer than with other media. Perfusate medium did not influence venous flow rates or absorption of Met. Effect of restricting venous flow on absorption of Lys and Met was evaluated in two calves. Flow was alternately controlled (6.5 ml/min) or allowed to flow freely (mean = 12.2 ml/min). Restricting flow decreased steady-state absorption. Light and scanning microscopy indicated maintenance of mucosal tissue integrity throughout 8 h of anesthesia. Results demonstrate validity of the in situ technique to study nutrient absorption in the young bovine.