Dynamic gut microbiome changes following regional intestinal lymphatic obstruction in primates.

The pathogenesis of inflammatory bowel disease (IBD) has been linked with lymphostasis, but whether and how lymphatic obstruction might disturb the intestinal microbiome in the setting of Crohn's Disease (CD) is currently unknown. We employed a new model of CD in African Green monkeys, termed 'ATLAS' (African green monkey truncation of lymphatics with obstruction and sclerosis), to evaluate how gut lymphatic obstruction alters the intestinal microbiome at 7, 21 and 61 days. Remarkable changes in several microbial sub- groupings within the gut microbiome were observed at 7 days post-ATLAS compared to controls including increased abundance of Prevotellaceae and Bacteroidetes-Prevotella-Porphyromonas (BPP), which may contribute to disease activity in this model of gut injury. To the best of our knowledge, these findings represent the first report linking lymphatic structural/gut functional changes with alterations in the gut microbiome as they may relate to the pathophysiology of CD.

Parenteral selenium and vitamin E supplementation of weaned beef calves.

BACKGROUND: The weaning process in beef calves elicits physiological stress responses that can negatively affect performance and health. Supplementation of vitamins and minerals, particularly selenium (Se) and vitamin E, might be warranted. HYPOTHESIS: That parenteral administration of Se or vitamin E would correct or prevent deficiencies of these nutrients in postweaned beef calves during a backgrounding period (42 days after weaning). ANIMALS: One hundred and forty-four weaned steers (200-250 kg) in 3 trials. METHODS: One trial was conducted with cattle on a dry lot fed a total mixed ration and 2 trials were conducted on cattle on pasture. Selenium was administered at 0.05 mg/kg BW (SC) and vitamin E was administered at 1500 IU on days 0 and 28 (SC). RESULTS: A treatment effect attributable to Se or vitamin E supplementation on average daily gain was not detected in any trials. Parenteral supplementation with Se on days 0 and 28 resulted in higher serum Se concentrations as compared with controls on day 42. Parenteral supplementation with Se on days 0 and 28 improved Se status from marginal to adequate in 1 trial. Parenteral supplementation with vitamin E did not improve serum vitamin E concentrations in any experiment. CONCLUSIONS AND CLINICAL IMPORTANCE: Supplementation with vitamin E or Se or a combination of both did not have a significant effect on calf performance during the 42-day backgrounding period.

Influence of finishing systems on hydrophilic and lipophilic oxygen radical absorbance capacity (ORAC) in beef.

The aim of this research was to: (1) develop a reliable extraction procedure and assay to determine antioxidant activity in meat products, and (2) assess the effect of beef finishing system (forage-finished: alfalfa, pearl millet or mixed pastures vs. concentrate-finished) on longissimus muscle antioxidant activity. The effect of extraction method (ethanol concentration and extraction time), protein removal, and sample preparation method (pulverization or freeze drying) were first evaluated to develop an antioxidant assay for meat products. Beef extracts prepared with low ethanol concentrations (20%) demonstrated higher hydrophilic ORAC. Protein removal prior to extraction reduced hydrophilic ORAC values. Sample preparation method influenced both hydrophilic and lipophilic ORAC, with pulverized samples containing higher hydrophilic and lipophilic ORAC values. Beef cattle finishing system (Forage: alfalfa, pearl millet, or natural pasture vs. concentrates) had little impact on muscle hydrophilic ORAC, but muscle from forage finished beef contained greater lipophilic ORAC. In addition, broiling of steaks reduced hydrophilic ORAC.

Evaluating the accuracy of a three-term pencil beam algorithm in heterogeneous media.

The goal of this work was to evaluate the accuracy of our in-house analytical dose calculation code against MCNPX data in heterogeneous phantoms. The analytical model utilizes a pencil beam model based on Fermi-Eyges theory to account for multiple Coulomb scattering and a least-squares fit to Monte Carlo data to account for nonelastic nuclear interactions as well as any remaining, uncharacterized scatter (the 'nuclear halo'). The model characterized dose accurately (up to 1% of maximum dose in broad fields (4 × 4 cm2 and 10 × 10 cm2) and up to 0.01% in a narrow field (0.1 × 0.1 cm2) fit to MCNPX data). The accuracy of the model was benchmarked in three types of stylized phantoms: (1) homogeneous, (2) laterally infinite slab heterogeneities, and (3) laterally finite slab heterogeneities. Results from homogeneous phantoms and laterally infinite slab heterogeneities showed high levels of accuracy (>98% of points within 2% or 0.1 cm distance-to-agreement (DTA)). However, because range straggling and secondary particle production were not included in our model, central-axis dose differences of 2-4% were observed in laterally infinite slab heterogeneities when compared to Monte Carlo dose. In the presence of laterally finite slab heterogeneities, the analytical model resulted in lower pass rates (>96% of points within 2% or 0.1 cm DTA), which was attributed to the use of the central-axis approximation.

Biophysical characterization of one-, two-, and three-tandem repeats of human mucin (muc-1) protein core.

Until recently mucin tandem repeat protein cores were believed to exist in random-coil conformations and to attain structure solely by the addition of carbohydrates to serine and threonine residues. Matsushima et al. (Proteins Struct. Funct. Genet., 7: 125-155, 1990) recently proposed a model of the secondary structure of proline rich tandem repeat proteins that has challenged this idea, especially for the case of the human polymorphic epithelial mucin encoded by the muc-1 gene. We report here results of structural analyses of the muc-1 protein core by using synthetic peptide analogues. Synthetic peptides were prepared to correspond to one-, two-, and three-tandem repeats of muc-1. Results of one- and two-dimensional 1H NMR correlation spectroscopy on these peptides confirm that the muc-1 protein core is not a random-coil secondary structure. Long-lived amide protons are protected in D2O, and increasing spectral complexity in the region of the beta-protons of Asp2 and His 15 reveals that structural changes are occurring as the number of repeats increases. The greatest changes occur when the number of repeats increases from one to two. These results are supported by the reactivity of a panel of monoclonal antibodies raised against tumor associated muc-1 with these synthetic peptides in enzyme-linked immunosorbent assay. The primary immunodominant mucin epitope, PDTRP, does not appear to attain a native conformation in the single repeat peptide (20 amino acids, starting with P), but is expressed on peptides with multiple repeats. Intrinsic viscosity measurements of the peptide containing three repeats indicate that an ordered structure present in solution is rod shaped. The circular dichroism spectrum of the same peptide is dominated by proline in the trans conformation. These results are all consistent with the prediction that the muc-1 tandem repeat polypeptide core forms a polyproline beta-turn helix.

Humoral immunity against a tandem repeat epitope of human mucin MUC-1 in sera from breast, pancreatic, and colon cancer patients.

Using synthetic peptides 60,80, and 105 residues long, corresponding to 3, 4, and 5.25 tandem repeats of human mucin MUC-1 protein core, as antigens in a solid-phase enzyme-linked immunosorbent assay, we screened sera from 24 breast cancer patients, 10 colon cancer patients, and 12 pancreatic cancer patients, at various stages of disease, for the presence of mucin-specific antibodies. The 105-residue peptide was superior in allowing detection of high levels of anti-mucin antibodies in 10.9% of sera in each cancer group. Another 4.3% showed intermediate reactivity. Lower levels of detection were achieved with the 80-residue peptide, and no specific reactivity was detectable with the 60-residue peptide. Anti-mucin antibodies were previously undetectable when this assay was performed with purified whole mucin or short synthetic peptides. The presence or absence of antibody did not correlate with the levels of circulating mucin or stage of disease. One highly reactive serum sample was used to identify more precisely the epitope on the long synthetic peptide to which the reactivity was directed. The reactivity of this serum specific for the 105-residue peptide was blocked by a 9-residue peptide from the NH2-terminal region of the 20-residue tandem repeat containing the previously identified immunogenic epitope APDTRP. Another 9-residue mucin peptide, from the COOH-terminal region of the tandem repeat which does not contain the APDTRP epitope, had no effect. All the mucin-specific reactivity was found to be of the IgM isotype, indicating a helper T-cell-independent response, unusual for an antibody against a peptide epitope, but not unexpected for tandemly repeated epitopes.

N-acetylgalactosamine glycosylation of MUC1 tandem repeat peptides by pancreatic tumor cell extracts.

Synthetic peptides corresponding to the human mucin MUC1 tandem repeat domain (20 residues) were glycosylated in vitro by using UDP-N-[3H]acetyl-D-galactosamine (GalNAc) and lysates of pancreatic tumor cell lines. Results obtained with peptides of different lengths (from one to five repeats) suggest that increasing the number of tandem repeats has neither a positive nor a negative effect on the density of glycosylation along the MUC1 tandem repeat protein backbone. Purified glycopeptides were sequenced on a gas-phase sequencer, and glycosylated positions were determined by measuring the incorporated radioactivity in fractions collected following each round of Edman degradation. The results showed that two of three threonine residues on the MUC1 tandem repeat peptides were glycosylated by pancreatic tumor cell lysates at the following positons: GVTSAPDTRPAPGSTAPPAH (underlined T indicates position of GalNAc attachment). None of the serine residues were glycosylated. Determination of the mass of the glycopeptides by mass spectrometry confirmed that a maximum of two molecules of GalNAc were covalently linked to each 20-residue repeat unit in the peptides. The data presented here show that acceptor substrate specificity of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase detected in lysates of pancreatic and breast tumor cell lines is identical and is limited to some but not all threonines in the MUC1 tandem repeat peptide sequence. The influence of primary amino acid sequence on acceptor substrate activity was evaluated by using several peptides that contain single or double amino acid substitutions (relative to the native human MUC1 sequence). These included substitutions in the residues that were glycosylated and substitutions of the surrounding primary amino acid sequence. The results of these studies suggest that primary amino acid sequence, length, and relative position of the residue to be glycosylated dramatically affect the ability of peptides to serve as acceptor substrates for the UDP-GalNAc:polypeptide N-acetylgalatosaminyltransferase.

Design tools for proton therapy nozzles based on the double-scattering foil technique.

Proton therapy has been increasing over the past several years, with several new treatment facilities being built in Europe, Japan and the United States. In this work, analytical and Monte Carlo tools were combined to model the passively scattered neurosurgery treatment beamline of the Harvard Cyclotron Laboratory (Cambridge, MA). The predicted three-dimensional dose distributions agree with actual measurements to within 0.1 mm for all quantities considered in central-axis depth-dose curve and to within 2.1 mm for all quantities considered in the absorbed dose cross-field profile. The predicted neutron dose equivalent per therapeutic absorbed dose, H/D, was calculated at various locations representing clinically significant anatomical sites. Under typical treatment conditions, the average ratio of predicted-to-measured H/D is 1.8 in the gonadal region (50 cm from isocentre) and 3.4 in the thyroid region (21 cm from isocentre). The global ratio of predicted-to-measured H/D is 2.6.

Structure-based design of peptides that recognize the CD4 binding domain of HIV-1 gp120.

DESIGN: Envelope protein-specific antiviral peptides, called mucibodies, that can specifically recognize and bind to the surface unit protein gp120 of HIV-1 were designed. The initial mucibody binding target was the V3 loop of HIV-1 gp120. Here, the gp120-CD4 binding domain was chosen as the site of mucibody binding. The CD4 binding domain of gp120 is known to be a conformational epitope and is involved in the earliest events of viral entry into many cells. METHODS: The design of the mucibody antivirals was based on previous observations that antibody complementarity determining regions (CDR) are generally similar to the repeating loops or knob structures found in the 20-residue tandem repeat domain of human mucin MUC1. The heavy chain CDR3 from the bacteriophage display antibody b12 was used to construct two mucibodies, b12-CDR1 and b12-26. RESULTS: Peptides corresponding to three tandem repeats were shown to bind directly to the CD4 binding domain of HIV-1 gp120 in a solid-phase enzyme-linked immunosorbent assay. These mucibody peptides also disrupted the gp120-CD4 interaction in a solution-phase inhibition assay. Finally, mucibodies neutralized primary and laboratory macrophage-tropic isolates of HIV-1. CONCLUSIONS: There is a potential for medical use of these peptides as topical vaginal microbicides in preventing HIV-1 transmission during sexual contact. These results also suggest that multivalent, non-immunogenic binding proteins of virtually any specificity could be constructed for use in therapeutic applications involving infectious diseases and immune system dysfunction.

Presentation of HIV V3 loop epitopes for enhanced antigenicity, immunogenicity and diagnostic potential.

OBJECTIVE: To evaluate the immunological properties of a panel of human mucin MUC1/HIV V3 loop chimeras. DESIGN: The immunodominant epitope of MUC1 (APDTR) was found to be structurally isomorphous with the tip of the principle neutralizing determinant (PND) of HIV-1 (MN) (GPGRA). A panel of 120 residue, six tandem repeat (TR) and 60 residue, three TR chimeric antigens were constructed in which the repeating MUC1 epitope is replaced by HIV-1 PND. Each 20 residue TR contains one PND epitope. The PND of HIV-1 is presented in the native beta-turn conformation at the crest of each repeating knob structure of the mucin-like protein. METHODS: The antigenicity of the chimeric antigens were compared using enzyme-linked immunosorbent assay (ELISA) and HIV-infected patient sera. Structural effects of antibody-antigen interactions were determined using surface plasmon resonance, with human monoclonal antibodies, chimeric antigens and the cyclic and linear V3 loops. Immunogenicity of three versus six TR was measured in mice. RESULTS: Nine residues of the HIV PND substituted into the mucin backbone were equivalent to the 36 residue cyclic V3 loop in ELISA. The 120 residue antigens induced high titer, immunoglobulin (Ig) M and IgG, and HIV-specific antibodies in mice. CONCLUSIONS: MUC1/V3 chimeras efficiently detect HIV-specific antibodies in patient sera. Multivalent presentation of the PND is advantageous for higher affinity antibody-antigen interactions and for inducing HIV-specific IgM and IgG antibodies.

Isolation and characterization of a proteoglycan variant from human aorta exhibiting a marked affinity for low density lipoprotein and demonstration of its enhanced expression in atherosclerotic plaques.

Proteoglycans (PG) are implicated in the pathophysiology of atherosclerosis due to their ability to complex with plasma low density lipoproteins (LDL). Studies were conducted to determine whether human aorta contains PG subclasses that exhibit enhanced LDL binding ability. PG were isolated from normal and atherosclerotic aortas by a combination of dissociative extraction and ion-exchange chromatography. The PG were further subfractionated on an LDL affinity column based on their binding affinity to LDL. Two PG fractions exhibiting high-affinity binding to LDL, as evidenced by their elution at 1.0 and 1.5 M NaCl, respectively, were isolated from both normal and atherosclerotic tissue. Compared with normal tissue, atherosclerotic tissue showed a twofold increase in the high-affinity PG that eluted at 1.5 M NaCl. Gel filtration of the high-affinity PG from normal tissue yielded two peaks (nPG2 and nPG3), while the high-affinity PG from plaque tissue was resolved into three peaks (pPG1, pPG2, and pPG3). pPG1 eluted at the void volume of the column, indicating that it was of very large molecular size. The hydrodynamic size of pPG2 was larger than that of the corresponding nPG2 (Kav = 0.44 versus 0.51), while pPG3 had the same hydrodynamic size as nPG3 (Kav = 0.86). The high-affinity PG subfractions from normal aorta contained varying proportions of chondroitin sulfates, dermatan sulfates, and heparan sulfate. In contrast, the PG subfractions from plaque tissue contained predominantly chondroitin sulfates and heparan sulfate. In vitro complexes of LDL and the high-affinity PG fractions from normal aorta and plaque tissue stimulated cholesteryl ester synthesis in human monocyte-derived macrophages. However, the LDL-plaque PG complex was significantly more potent than the LDL-normal aorta PG complex in this respect. These results indicate that PG subclasses with enhanced binding affinity to LDL occur in the normal human aorta and that their concentration increases significantly in atherosclerotic lesions. In addition, the high-affinity PG in plaque tissue have altered characteristics and increased ability to stimulate LDL-mediated cholesterol ester synthesis in macrophages. This could lead to increased lipid deposition during atherogenesis.

Structure and self assembly of a retrovirus (FeLV) proline rich neutralization domain.

The 60 amino acid proline-rich neutralization domain of the external surface unit glycoprotein of feline leukemia virus was chemically synthesized in total and in fragments. We examined the ability of these retroviral peptides to form ordered conformations using 1H-NMR, circular dichroism spectroscopy, and intrinsic viscosity measurements. One dimensional nuclear magnetic resonance spectroscopy revealed that the 60 amino acid peptide could form a stable, folded structure that was long-lived, as shown by the ability to protect amide-protons in D20. Peptides corresponding to the N-terminal 42, N-terminal 20 amino acids, and middle 20 amino acid sections could also form stable structures. The C-terminal segment did not protect any protons in D20. Interestingly, self assembly of the N-terminal 42 and C-terminal 16 amino acid peptides into a structure very close to that of the 60 amino acid domain was observed. The circular dichroism results reveals a large negative cotton effect at 198 nm that is characteristic of the proline-rich beta-turn helixes which consist predominantly of trans-proline. The intrinsic viscosity results suggest a non-random coil structure that is rod shaped. Our conclusion is that PRN60 forms a beta-turn helix and that this region of FeLV-gp70 is a separate folding domain of the retroviral surface unit glycoprotein. The unique conformational properties of PRN60 and its critical role as the predominant target for neutralizing antibody responses suggest that this peptide is a reasonable candidate for producing a synthetic peptide vaccine for FeLV.

Structure of a tumor associated antigen containing a tandemly repeated immunodominant epitope.

Human mucins are T or S glycosylated tandem repeat proteins. In breast cancer, mucins become under or unglycosylated. Two-dimensional nuclear magnetic resonance experiments are performed on chemically synthesized mucin tandem repeat polypeptides, (PDTRPAPGST-APPAHGVTSA)n the unglycosylated form for n=1,3 where (APDTR) constitutes the antigenic sites for the antibodies isolated form the tumors in the breast cancer patients. These studies demonstrate how the tandem repeats assemble in space giving rise to the overall tertiary structure, and the local structure and presentation of the antigenic site(APDTR) at the junction of two neighboring repeats. The NMR data reveal repeating knob-like structures connected by extended spacers. The knobs protrude away from the long-axis of Muc-1 and the predominant antigenic site (APDTR) forms the accessible tip of the knob. Multiple tandem repeats enhance the rigidity and presentation of the knob-like structures.

Evaluation of humoral immune responses in cattle grazing endophyte-infected or endophyte-free fescue.

Anecdotal reports suggest cattle with fescue toxicosis may not respond to vaccination and thus, experience increased incidence of Bovine Respiratory Disease Complex (BRDC) when shipped to feedlots. Fescue toxicosis causes hypoprolactemia in cattle. Hypoprolactemia decreases humoral immune responses in mice. Therefore, a study was conducted to compare the magnitude of primary and secondary humoral immune responses against specific antigens in cattle grazing endophyte-infected or endophyte-free fescue. Angus steers were blocked by weight and allocated into four groups. Two groups grazed endophyte-infected (EI) fescue and the other two groups grazed endophyte-free (EF) fescue. All steers were injected IM on d 0 and 21 with lysozyme without adjuvant and concanavalin. A (Con A) with sheep red blood cells (SRBC) in incomplete adjuvant of Freund. Steers were bled on days 0, 21 and 35 post-vaccination. Average daily gains (ADG), alkaline phosphatase (ALP) activity, cholesterol concentrations, rectal temperatures, and serum prolactin concentrations were measured to confirm fescue toxicosis in steers grazing EI fescue. Antibodies to Con A and SRBC were determined by ELISA and hemagglutination assay, respectively. The ADG were decreased for the EI group during the first month. Rectal temperature were elevated and serum prolactin concentrations were decreased in the EI group. Cholesterol and ALP concentrations also were decreased in the EI group. Primary and secondary immune responses against Con A tended to be increased and were increased against SRBC in the EI group. Antibodies against lysozyme were not induced in either group. In conclusion, cattle grazing EI fescue mounted similar humoral immune responses to vaccination, despite hypoprolactemia, as cattle grazing EF fescue. Increases in bovine respiratory disease in cattle maintained on EI fescue probably is not associated with lack of humoral immune response to vaccination protocols as a result of fescue toxicosis.

Overestimation of hemoglobin in a patient with an IgA-kappa monoclonal gammopathy.

A patient with multiple myeloma had an automated blood count performed on a Coulter STK-S counter that repeatedly failed internal limits for both mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. The calculated hematocrit agreed with a spun hematocrit, suggesting that the hemoglobin concentration was being overestimated by the automated counter. Measurement of the plasma hemoglobin concentration of the sample, which showed no visible hemolysis, gave a hemoglobin concentration of 32 g/L on the STK-S analyzer. Correction of the whole blood hemoglobin using the plasma hemoglobin gave a value consistent with the hematocrit. The corrected mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration values were within standard limits. This patient's paraprotein was characterized as IgA-kappa and was present at a concentration of 61 g/L. The hemoglobin concentration measured on whole blood by Sysmex NE 8000 and Technicon H*1E autoanalyzers agreed reasonably well with the corrected result from the STK-S.

Vesicouterine fistula after cesarean section. A case report.

A 27-year-old woman, gravida 3, para 2002, underwent repeat cesarean section and inadvertent cystotomy, with subsequent development of a vesicouterine fistula. The fistula did not resolve with prolonged bladder catheterization and required total abdominal hysterectomy with resection of the fistula tract and primary closure for definitive treatment.

Effects of intravenous infusion of high levels of potassium and sodium on mineral metabolism in sheep.

A metabolism trial was conducted with 18 crossbred (Finn x Dorset x Suffolk) wethers, fitted with indwelling jugular catheters and abomasal and ileal cannulas, to determine the effects of high levels of i.v.-infused K and Na on mineral metabolism. The wethers (40 kg) were fed 800 g daily of a 60% concentrate diet in two equal portions at 0800 and 1900. Six wethers were infused randomly with 19 g K+, six with 10.6 g Na+ (chloride salts) and the other six with physiological saline solution (1.2 g Na) per day. Potassium chloride or NaCl infusion had no effects on apparent absorption, retention, flow or partial absorption of Mg, Ca and P in the digestive tract compared with physiological saline infusion. With all treatments, Mg and Ca were absorbed proximal to the abomasal cannula. Magnesium was secreted into, whereas Ca and P were absorbed from, the small intestine. Phosphorus was secreted both in the stomach and large intestinal regions of the digestive tract. Major sites of K and Na absorption were the small and large intestines, respectively. Infusion of K increased (P less than .05) retention of K compared with Na infusion. Infusion of Na increased (P less than .05) excretion and retention of Na compared with K infusion. Serum minerals were not changed by K or Na infusion compared with saline. The results of this experiment indicate that the depressing effects of K on Mg absorption are not attributable to high levels of absorbed K, but rather to K present in the digestive tract prior to the small intestine.

Utilization of phosphorus and certain other minerals from swine waste and broiler litter.

Two trials were conducted with 15 wethers surgically equipped with duodenal and ileal cannulas to study the utilization of P, Ca, Mg, Cu, Fe and Zn from swine waste and broiler litter. For each trial, animals were fed a low-P basal diet until serum inorganic P averaged 5.5 mg/dl; then they were allotted at random to the following 50% DM ensiled diets: low-P basal, basal + swine waste, basal + broiler litter, basal + dicalcium phosphate and basal + soybean meal. Each trial consisted of a 7-d preliminary period, a 7-d collection of feces and urine and 6-d sampling of duodenal and ileal digesta and feces. Apparent P absorption was not different (P greater than .05) between sheep fed waste-supplemented diets (37%) and those fed the conventionally supplemented diets (28%). Phosphorus absorption, calculated by difference, tended (P less than .1) to be higher from the waste supplements (59%) than from dicalcium phosphate and soybean meal (37%). Less (P less than .05) Ca was absorbed from the waste diets (.62 g/d) than from the conventional diets (1.28 g/d). More (P less than .05) Cu (mg/d) was absorbed from the waste diets, but no difference was found when absorption was expressed as percentage of intake. Broiler litter and swine waste were good sources of available P and Mg for ruminants.

Utilization of magnesium and other macrominerals in sheep supplemented with different readily-fermentable carbohydrates.

Sheep were used to determine the effects of dietary supplementation with readily-fermentable carbohydrates on Mg, Ca, K and P utilization. In each of two metabolism trials, 15 mature, crossbred wethers (average weight, 49.2 kg) were allotted to five dietary treatments consisting of 800 g/d of orchardgrass (Dactylis glomerata, L.) hay alone, or supplemented with 450 g/d of either glucose, sucrose, lactose or starch. Each trial consisted of a 5-d adjustment period, a 10-d preliminary period and a 10-d collection period. Compared with wethers fed hay alone, supplementation with each kind of carbohydrate to the diet decreased (P less than .05) fecal Mg excretion and increased (P less than .05) apparent absorption and retention of Mg. Apparent absorption of the Ca was lower (P less than .05) in wethers fed lactose and tended to be decreased by glucose, sucrose and starch supplementation. Calcium retention was lower (P less than .05) in wethers fed sucrose and lactose, compared with those fed hay alone. All types of supplementary carbohydrates depressed (P less than .05) apparent absorption and urinary excretion of K. Serum Mg and Ca were not affected and serum K was depressed (P less than .05) by carbohydrate supplementation. Ruminal fluid pH was decreased (P less than .05) by glucose and lactose supplementation, and addition of these carbohydrates tended to decrease molar proportions of acetate and increase those of propionate and butyrate, compared with sheep fed hay alone. Sucrose addition decreased (P less than .05) acetate and increased (P less than .05) butyrate molar proportions in the ruminal fluid.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of supplemented magnesium from magnesium oxide or dolomitic limestone upon digestion and absorption of minerals in sheep.

Three metabolism trials were conducted with six lambs fitted with abomasal and ileal cannulae to determine the site of supplemental Mg absorption when supplied as dolomitic limestone or magnesium oxide. Magnesium intake was 1.09 g/d for the lambs fed the control diet and 2.21 and 2.10 g for those fed the dolomitic limestone and magnesium oxide supplemented diets, respectively. Irrespective of its source, the major site of Mg absorption was the preintestinal region. Magnesium was secreted in the small intestine in lambs fed all three diets. Magnesium supplementation resulted in an increase (P less than .05) in preintestinal Mg absorption, with magnesium oxide having the highest value. Supplementation had no effect (P greater than .05) on secretion or absorption of Mg from the small and large intestines or total retention of Mg. With all three diets, the major site of Ca absorption was the stomach region with additional absorption taking place in the large intestine. Calcium was secreted in the small intestine of lambs fed all treatments. Generally, K, Na and P were secreted in the preintestinal region and absorbed from the small and large intestines. The major site of K and P absorption appeared to be the small intestine and for Na, the large intestine. Serum minerals were unchanged (P greater than .05) due to treatment, but the levels of Mg, K and inorganic P tended to be higher in lambs supplemented with either of the Mg sources. It appears that the greater utilization of Mg from magnesium oxide vs dolomitic limestone is due to greater absorption of Mg from the forestomach region.