Direct fed microbial supplementation repartitions host energy to the immune system.

Direct fed microbials and probiotics are used to promote health in livestock and poultry; however, their mechanism of action is still poorly understood. We previously reported that direct fed microbial supplementation in young broilers reduced ileal respiration without changing whole-body energy expenditure. The current studies were conducted to further investigate the effects of a direct fed microbial on energy metabolism in different tissues of broilers. One hundred ninety-two 1-d-old broiler chicks (16 chicks/pen) were randomly assigned to 2 dietary groups: standard control starter diet (CSD) and CSD plus direct fed microbial (DFMD; 0.3%) with 6 pens/treatment. Body weight, feed consumption, whole-body energy expenditure, organ mass, tissue respiration rates, and peripheral blood mononuclear cell (PBMC) ATP concentrations were measured to estimate changes in energy metabolism. No differences in whole body energy expenditure or BW gain were observed; however, decreased ileal O(2) respiration (P < 0.05) was measured in DFMD fed broilers. In contrast, the respiration rate of the thymus in those broilers was increased (P < 0.05). The PBMC from DFMD fed broilers had increased ATP concentrations and exhibited increased ATP turnover (P < 0.01). To determine if the increased energy consumption by PBMC corresponded with an altered immune response, broilers were immunized with sheep red blood cells (SRBC) and assayed for differences in their humoral response. The DFMD-fed broilers had a faster rate of antigen specific IgG production (P < 0.05) and an increase in total IgA (P < 0.05). Collectively, these data indicate that supplementation with the direct fed microbial used in this study resulted in energy re-partitioning to the immune system and an increase in antibody production independent of changes in whole body metabolism or growth performance.

Direct-fed microbial PrimaLac and salinomycin modulate whole-body and intestinal oxygen consumption and intestinal mucosal cytokine production in the broiler chick.

The current study investigated whole-body O2 consumption, intestinal O2 consumption, and intestinal inflammation status through mucosal cytokine production on broiler chicks fed the direct-fed microbial PrimaLac. One hundred twenty 1-d-old broiler chicks were randomly assigned to 1 of 3 experimental diets: standard starter diet (control), standard starter diet with added salinomycin (SAL), and standard starter diet with added PrimaLac (DFM). Birds were housed in 2 separate rooms, the control and SAL treatments in one room and the DFM in another. Intact ileal and cecal samples were collected on d 19, 20, and 21 after measuring whole-body O2 consumption using indirect calorimetry. The O2 up-take of ileal tissue was measured using an in vitro O2 monitor. Analysis of intestinal immune status of broilers was measured by the relative differences in mRNA of both pro- and antiinflammatory cytokines: interleukin-(IL) 1beta, IL-6, and IL-10 using real-time reverse transcription-PCR. Broilers exhibited a 6 to 16% decrease in whole-body energy expenditures and up to a 47% decrease (P<0.05) in ileal energy expenditures in the DFM group compared with other treatments. The reverse transcription-PCR data demonstrated that DFM consortium numerically altered both pro- and antiinflammatory cytokines within the ileum of 19-d posthatch broilers. These data suggest that direct-fed microbials like PrimaLac increase metabolic efficiency via changes in intestinal physiology and metabolism.

Ionophores have limited effects on jejunal glucose absorption and energy metabolism in mice.

Two experiments, Trial 1 (in vitro) and Trial 2 (in vivo), were conducted to examine the effects of ionophores, monensin, laidlomycin, and laidlomycin propionate on whole-animal O2 consumption, organ weights, jejunal glucose absorption, and O2 utilization, as well as growth, feed and water consumption, and feed efficiency. In Trial 1, 30 male Swiss-Webster mice, 8 wk old, were used to measure the in vitro effects of each of the ionophores at concentrations of 1.62 or 16.2 mM. Six combinations of three ionophores at two concentrations resulted in a total of eight treatments. All eight treatments were exposed to jejunal rings from a single mouse for a total of 30 observations per treatment. Jejunal rings were exposed to each ionophore treatment for 15 min. Laidlomycin propionate (16.2 mM) decreased (P < 0.02) glucose absorption, as estimated by H3-3-O-methyl glucose uptake compared with all other treatments, whereas laidlomycin propionate (1.62 mM) increased (P = 0.032) jejunal DM content compared with 16.2 mM laidlomycin propionate. In Trial 2, 40 5-wk-old mice were allotted into four treatments--control and 16.2 mM each of monensin, laidlomycin, and laidlomycin propionate--for a total of 10 observations per treatment. Ionophores were administered via the drinking water for 14 d. No ionophore treatment had any effect on whole-mouse O2 consumption. Monensin increased (P = 0.004) stomach size and decreased (P = 0.049) the efficiency of BW gain compared with controls. Laidlomycin propionate decreased (P = 0.032) the percentage of whole jejunum oxygen consumption due to oubain-sensitive respiration compared with control. The efficiency of intestinal glucose absorption was not changed due to treatment in either trial. Under the conditions of these studies, monensin, laidlomycin, and laidlomycin propionate had minimal and inconsistent effects on jejunal function and energy utilization in mice. This investigation suggests that changes in the energetic requirements of animals treated with ionophores are not an issue in animal production.

Administration of triiodothyronine and dopamine to broiler chicks increases growth, feed conversion and visceral organ mass.

The influences of triiodothyronine (T3) or dopamine (DA) administration on growth, feed conversion, and visceral weights in broiler chicks between the ages of 6 and 12 d posthatch were investigated. In Trial 1, six chicks at age 6 d were randomly administered one of the following treatments: 0.37, 0.74, 1.48, and 2.96 micromol T3/kg BW or 0.07, 0.14, 0.28, and 0.56 micromol DA/kg BW. Both T3 and DA were administered via intraperitoneal injections between the end of sternum and the ends of os pubis, with 0.9% saline as the excepient. In addition, two groups of six birds each were either not injected or injected with excepient only, as controls. Four replications were carried out with a total of 264 chicks. Heart weight as a percentage of feed-deprived body weight (FDBW) of the chicks injected with 2.96 micromol T3/kg BW was heavier than that of controls. Other variables measured were not significantly different between treatments. In trial 2, six chicks at age 6 d were randomly administered, one of the following treatments: 0.56, 1.12, 2.24, and 4.48 micromol T3/kg diet or 0.40, 0.80, 1.60, and 3.20 micromol DA/kg diet as well as a nonsupplemented control. Four replications were carried out with 216 chicks. The results in Trial 2 showed that the effects of T3 (X, micromol/kg diet) on body weight gain (Y1, g) and feed consumption (Y2, g) were linear (Y1 = 310 - 21.5X, R2 = 0.868, P < 0.001 and Y2 = 398 - 22.3X, R2 = 0.765, P < 0.001, respectively). The feed conversion ratio, the weight of liver, the weights of various intestinal segments, the lengths of the duodenum, jejunum and the ileum, as well as weight per centimeter jejunal length, gizzard weight as percentage of FDBW, and the duodenal length per kilogram FDBW all had linear responses (P < 0.05) to the level of dietary supplementation of T3. The effect of dietary supplementation of T3 on the heart weight was quadratic (Y16 = 2.58 + 0.89X - 0.17 X2, R2 = 0.526, P < 0.01). Similarly, the weights of pancreas and gizzard, the heart weight as a percentage of FDBW and the pancreas weight as a percentage of FDBW all had second-order curve responses. Dietary DA supplementation exerted no effect on the variables measured except that the regression of the heart weight as a percentage of FDBW on dietary DA supplementation (X1, micromol/kg diet) existed, namely, Z1 = 0.64 + 0.24 X1 - 0.23 X1(2) + 0.05 X1(3) (R2 = 0.868, P < 0.05).

In ovo peptide YY and epidermal growth factor administration and their effects on growth and yolk utilization in neonatal meat-type chickens (Gallus domesticus).

The effects of in ovo peptide YY (PYY) or epidermal growth factor (EGF) administration on chick growth, yolk absorption and yolk stalk function in posthatch (0-5 days) meat-type or broiler chicks were determined. At Day 18 of incubation, treated eggs were injected into the air cell with 100 microl of either PYY (Trial 1) or EGF (Trial 2) at a dosage of 600 microg/kg egg weight. Saline-treated control eggs were injected similarly with 0.9% saline. At hatch, 200 microl of (51)Cr-labeled microspheres were injected into chick yolk sacs. Epidermal growth factor increased ileal wet weight adjusted for body weight as well as ileal serosal dry matter. Body weight, feed consumption and excreta weight per bird, and relative weights of the yolk sac, intestine and liver were significantly affected by age of the chick in both trials. Relative radioactivity of the yolk sac, yolk stalk, blood, liver, and kidneys were affected by bird age in Trial 2; however, there were no significant effects due to PYY or EGF treatments on relative radioactivity of the tissues and organs examined. These data suggest that PYY and EGF had no effect on yolk absorption or yolk stalk function through 5 days in the posthatch chick.

Neuropeptide Y, peptide YY and aluminum in Alzheimer's disease: is there an etiological relationship?

Neuropeptide Y (NPY) and peptide YY (PYY) are members of the pancreatic polypeptide family which have a high degree of primary and tertiary structural homology. They function as neurotransmitters and humoral agents in central nervous system and gastrointestinal function. During the last two decades, NPY body fluid concentrations and NPY/PYY brain receptor numbers have been demonstrated to be altered during the course of Alzheimer's disease. Recent research has shown that both NPY and PYY may be involved in aluminum metabolism in animal models. A brief discussion of the structure, biological activity and possible involvement of these peptides in aluminum metabolism and Alzheimer's disease is contained herein.

Peptide YY administration decreases brain aluminum in the Ts65Dn Down syndrome mouse model.

We have previously reported the Ts65Dn (Ts) mouse has impaired intestinal absorptive function and amino acid metabolism. Peptide YY (PYY) has enhanced glucose absorption in mice and turkeys. Other studies have reported that persons with Down syndrome have increased intestinal absorption of aluminum. Alzheimer's-like lesions have been reported in Ts mice. Trial 1 of this study examined brain Al concentrations, plasma metabolites and intestinal metabolism of 40 control and 40 Ts mice administered 300microg PYY/kg body weight or 0.9% saline for 3d. Trial 2 examined nutrient digestibility of 12 C and 12 Ts given PYY or saline for 14d. In Trial 1, PYY lowered (p<0.05) the brain Al pool (mg/g FBW) in both C and Ts mice by 80% compared to saline. Ts mice had increased plasma NH3 (329 vs. 269 microM, p<0.05), decreased plasma glucose (7.4 vs. 8.4 mM, p<0.01), elevated apparent energetic efficiency of jejunal glucose uptake (p<0.01) and elevated brain Al pool (0.41 vs. 0.12 microg, p=0.06) compared to C mice. In Trial 2, PYY increased small intestinal density (mg/cm) 12% in both genotypes (p<0.05), but did not alter nutrient digestibility. Brain Al accretion and hyperammonemia are proposed risk factors for Alzheimer's disease (AD). Ts mice and PYY appear to be suitable models for the study of metabolic and neurological anomalies in Down syndrome and AD.

Spinal integration of antidromic mediated cutaneous vasodilation during dorsal spinal cord stimulation in the rat.

The purpose of this study was to determine the involvement of supraspinal centers and spinal synaptic integration in cutaneous vasodilation mediated by dorsal spinal cord stimulation (DCS). Laser Doppler flowmetry was used to assess cutaneous blood flow changes in the rat hindpaw during DCS with a unipolar ball electrode placed at the L2-L3 spinal level. Results demonstrated that transecting the spinal cord at the T10 spinal segment did not alter the DCS response while T13 spinal transection abolished the DCS-induced vasodilation. Inhibition of synaptic activity with topical application of muscimol (0.2 mM) on the dorsal surface of the spinal cord markedly attenuated the DCS response. In conclusion DCS-induced vasodilation involved synaptic integration but did not require input from rostral spinal sites or supraspinal areas.

Differential effects of fat and sucrose on body composition in A/J and C57BL/6 mice.

The C57BL/6 (B6) mouse is more sensitive to the effects of a high-fat diet than the A/J strain. The B6 mouse develops severe obesity, hyperglycemia, and hyperinsulinemia when fed this dietary regimen. This study was conducted to determine the effects of dietary fat and sucrose concentrations on body composition and intestinal sucrase (EC 3.2.1.48) and maltase (EC 3.2.1.20) activity in these two mouse strains. High-fat diets, regardless of sucrose content, resulted in significant weight gain, higher body fat, and lower body protein and water content in both strains of mice. The shift toward higher body fat and lower protein and water content was far greater in the B6 strain. Low-fat, high-sucrose diets resulted in lower body weight in both strains, as well as significantly greater body protein content in B6 mice. Analysis of intestinal sucrase showed that the enzyme was less active in B6 mice when the diet was high in sucrose. Both sucrase and maltase had lower activity in the presence of high dietary fat in both mouse strains. The percent reduction of intestinal enzyme activity due to dietary fat was similar in both strains. The B6 mouse exhibits disproportionate weight gain and altered body composition on a high-fat diet. This coupled with the reduced body weight and increased body protein on a low-fat, high-sucrose diet suggests that factors-relative to fat metabolism rather than sucrose metabolism are responsible for obesity.

Reevaluation of the Role of the Sympathetic Nervous System in Cutaneous Vasodilation during Dorsal Spinal Cord Stimulation: Are Multiple Mechanisms Active?

Objective. In addition to treatment of refractory chronic pain in patients with peripheral vascular disease, dorsal spinal cord stimulation (DCS) increases cutaneous blood flow to the extremities and may have a limb-saving effect. The purpose of this study was to examine the role of the sympathetic nervous system in the cutaneous vasodilation due to DCS. Methods. Male Sprague-Dawley rats were anesthetized with pentobarbital (60 mg/kg, i.p.). A unipolar ball electrode was placed on the left side of the exposed spinal cord at approximately the L1-L2 level. Blood flow was concurrently recorded from both hindpaw foot pads with laser Doppler flowmeters. Blood flow responses were assessed during 1 min of DCS (0.6 mA at 50 Hz, 0.2 msec pulse duration) at 10 min intervals. To determine the contribution of the sympathetic nervous system in the blood flow response to DCS, the role of ganglionic transmission, alpha-adrenergic receptors, beta-adrenergic receptors, and adrenal catecholamine secretion were investigated using adrenergic receptor antagonists. Results. Hexamethonium (10 mg/kg, i.v.), an autonomic ganglionic receptor antagonist, did not attenuate the cutaneous vasodilation during DCS. Phentolamine (3 mg/kg, i.v.), a nonselective alpha-adrenergic receptor antagonist, also did not attenuate the DCS-induced increase in peripheral cutaneous blood flow. On the other hand, prazosin (0.1 mg/kg, i.v.), a selective alpha-1-adrenergic receptor antagonist, attenuated the DCS response but this may, at least, be partly due to a vehicle effect. Propranolol (5 mg/kg, i.v.), a nonselective beta-adrenergic receptor antagonist, attenuated the DCS response while adrenal demedullation did not. Conclusion. Overall, our results show that DCS-induced vasodilation can occur through mechanisms that are independent of sympathetic outflow.

Role of nitric oxide in cutaneous blood flow increases in the rat hindpaw during dorsal column stimulation.

OBJECTIVE: Dorsal column stimulation (DCS) increases blood flow to the extremities and may produce a limb-saving effect in addition to treatment of refractory chronic pain in patients with peripheral vascular disease. The purpose of this study was to examine the importance of nitric oxide in cutaneous vasodilation caused by DCS. METHODS: Male Sprague-Dawley rats were anesthetized with pentobarbital (60 mg/kg, intraperitoneally). A unipolar ball electrode was placed on the left-side of the exposed spinal cord at approximately L1-L2. Blood flow was concurrently recorded from both hindpaw foot pads with laser doppler flowmeters. Blood flow responses were assessed during 1 minute of DCS (0.6 mA at 50 Hz, 0.2-ms pulse) at 10-minute intervals. Nitric oxide synthase was inhibited with NG-nitro-L-arginine methyl ester (L-NAME). Four groups of animals were examined. The first and second groups involved examination of the effects of DCS after 2 and 10 mg/kg L-NAME, respectively. In the third group, the effect of another nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (10 mg/kg), was examined on the responses to DCS. The fourth group of animals entailed comparison of the effects of DCS under control conditions, after the nicotinic receptor antagonist, hexamethonium (10 mg/kg), and during the combined presence of hexamethonium and L-NAME (10 mg/kg). RESULTS: L-NAME markedly attenuated the cutaneous blood flow increases caused by DCS at doses of 2 or 10 mg/kg. Similarly, NG-monomethyl-L-arginine also attenuated the DCS response. Hexamethonium did not affect the cutaneous vasodilation caused by DCS. After hexamethonium, L-NAME no longer attenuated the DCS response. CONCLUSION: Our results demonstrated that nitric oxide played a significant role in producing the DCS-induced increase in rat cutaneous hindpaw blood flow. The involvement of nitric oxide does not require the presence of autonomic efferent function; however, ganglionic blockade may unmask a mechanism for vasodilation during DCS that is independent of nitric oxide release.

Cutaneous vasodilation during dorsal column stimulation is mediated by dorsal roots and CGRP.

Dorsal column stimulation (DCS) is used clinically to provide pain relief from peripheral vascular disease and has the benefit of increasing cutaneous blood flow to the affected lower extremities. The purpose of this study was to examine the role of dorsal roots, calcitonin gene-related peptide (CGRP), and substance P in the cutaneous vasodilation induced by DCS. Male rats were anesthetized with pentobarbital sodium (60 mg/kg ip). A unipolar ball electrode was placed unilaterally on the spinal cord at the L1-L2 spinal segment. Blood flow was recorded in each hindpaw foot pad with laser Doppler flowmeters. Blood flow responses were assessed during 1 min of DCS (either 0.2 mA subdural or 0.6 mA epidural at 50 Hz, 0.2-ms pulse duration). Dorsal rhizotomy of L3-L5 (n = 5) abolished the cutaneous vasodilation to subdural DCS, whereas removal of T10-T12 (n = 5) and T13-L2 dorsal roots (n = 5) did not attenuate the DCS-induced vasodilation. The CGRP antagonist, CGRP-(8-37) (2.6 mg/kg iv, n = 7), eliminated the epidural DCS-induced vasodilation, whereas the substance P receptor antagonist, CP-96345 (1 mg/kg iv, n = 6), had no effect. In summary, L3-L5 dorsal roots and CGRP are essential for the DCS-induced vasodilation. We propose that DCS antidromically activates afferent fibers in the dorsal roots, thus causing peripheral release of CGRP, which produces cutaneous vasodilation.

Jejunal glucose uptake and oxygen consumption in turkey poults selected for rapid growth.

Two lines of turkey poults, one selected for rapid growth at 16 wk of age (F line) and the other a randombred control line (RBC2) were used to investigate the effect of selection for rapid growth on jejunal O2 consumption and glucose transport as well as whole-body O2 consumption. All trials used unsexed poults and were designed as a randomized complete block with day and line as independent variables. In Trial 1, 120 turkey poults, fed a standard starter ration (25.5% CP), were used to examine the effect of selection on feed intake, body weight gain, and efficiency from hatching (Day 0) to 13 d of age. At Day 14, 36 of 60 birds from each line were killed to measure intestinal length and weight and jejunal O2 consumption after 18 h of feed deprivation. Compared with the RBC2 line, the F line had relatively shorter but heavier small intestinal segments when adjusted by 18 h feed-deprived body weight (FBW; P < 0.001). The F line consumed more O2 over the entire jejunum adjusted to FBW than RBC2 line (43.8 vs 34.6 nmol O2/min.g FBW; P < 0.001). Jejunal ouabain- and cycloheximide-sensitive O2 consumption were greater (P < 0.05) in the F line. In Trial 2, 16 14-d-old poults from each line were used to measure in vitro jejunal glucose transport rate. There was no difference in glucose transport of the jejunum (nanomoles per minute per gram of FBW) between the lines. In Trial 3, 20 poults from each line were used to measure whole-body O2 consumption at 7 to 10 d of age. The F and RBC2 lines had similar whole-body O2 consumption rate per gram of FBW. These data suggest that selection of turkeys for rapid growth at 16 wk of age did not increase efficiency of jejunal glucose uptake in 14-d-old turkey poults.

Cutaneous blood flow increases in the rat hindpaw during dorsal column stimulation.

The purpose of this study was to determine the optimal stimulation site and parameters that result in the greatest changes in cutaneous blood flow during dorsal column stimulation (DCS). Laser Doppler flowmetry was used to assess cutaneous blood flow changes in both rat hindpaws during DCS with a unipolar ball electrode. We found that stimulating the dorsal column at the L2 spinal segment at 0.6 mA at either 25 or 50 Hz with a pulse duration of 0.2 ms resulted in the largest cutaneous blood flow increases in the rat hindpaw. In addition, the DCS response appeared to be limited primarily to the hindpaw ipsilateral to the site of DCS.

T cell localization in rabbit models of acute and chronic experimental hypersensitivity pneumonitis.

Cell-mediated hypersensitivity has been increasingly implicated in immunologic diseases of the lung, including hypersensitivity pneumonitis (HSP) (extrinsic allergic alveolitis). We used a T cell-specific monoclonal antibody (L11/135) to localize T cells in the parenchyma and bronchus-associated lymphoid tissue of ethanol-fixed, paraffin-embedded lung sections in rabbit models of experimental HSP to define further their possible role in pathogenesis. T cells appeared within 4 hours in early lesions of rabbit models of acute HSP and heavily infiltrated alveolitis lesions at 8 and 24 hours after aerosol challenge. T cells were also present in lesions of rabbit models with chronic alveolitis and occurred peripherally in granulomas. Variable aggregate and follicular forms of bronchus-associated lymphoid tissue rich in T cells occurred in both experimental and control animals. Our findings document early and continuing presence of T cells in lesions in rabbit models of experimental HSP.