Vertical migratory behavior of the infective third-stage larvae of Oesophagostomum dentatum.

The vertical migratory behavior of third-stage infective larvae (L3i) of Oesophagostomum dentatum was investigated using upright truncated agarose cones and equivalent conical depressions in agarose. Geotactic response varied with the age of the infective larvae. Four-day-old L3i showed no preference for the sloping surfaces of either indented or upright cones, while the 8-day-old L3i showed a positive geotactic reaction, migrating down the sloping surface of the depressions.

Amphids: the neuronal ultrastructure of macrocyclic-lactone-resistant Haemonchus contortus.

The development of anthelmintic resistance by nematode parasites is a growing problem for veterinarians and producers. The intensive use of the macrocyclic lactones for the treatment of a variety of parasitic diseases has hastened the development of resistance to this family of parasiticides among sheep, goats and cattle. As a result, resistance to ivermectin, moxidectin and doramectin by Haemonchus contortus has been documented throughout the world. While the exact sites of action of the macrocyclic lactones remain incompletely known, a critical point of entry for these drugs may be the terminally exposed sensory major neurons located in the cephalic end of the worms. These neurons, called amphidial neurons, are located in a pair of channels, the amphids, on either side of the pharynx and are exposed to the external environment via pores at the anterior tip of the worm. Through these neurons, important chemical and thermal cues are gathered by the parasite. Examination of serial electron micrographs of ivermectin-susceptible and ivermectin-resistant H. contortus allows for comparison of neuronal structure, arrangement of neurons within the amphidial channel, and distance of the tip of the dendritic processes to the amphidial pore. The latter of these characteristics provides a useful means by which to compare the association between the neurons and the external environment of the worm. Comparison of parental laboratory strains of ivermectin-susceptible H. contortus with related selected, ivermectin-resistant strains and with a wild-type ivermectin-susceptible field strain of H. contortus from Louisiana reveals that the ivermectin-resistant worms examined have markedly shorter sensory cilia than their ivermectin-susceptible parental counterparts. Additionally, the amphidial neurons of ivermectin-resistant worms are characterized by generalized degeneration and loss of detail, whereas other neurons outside of the channels, such as the labial and cephalic neurons, are normal in structure. Similar degeneration was also observed in field strains of doramectin-resistant H. contortus collected from a New Jersey alpaca heard. These findings raise a number of questions regarding the relationship between amphidial structure and macrocyclic lactone resistance as well as the role of amphids as a means of entry for these molecules. While shortened amphidial sensilla are associated with ivermectin resistance, it remains unclear if such a structural modification facilitates survival of nematodes exposed to the macrocyclic lactones.

Effect of glucose on antipsychotic drug-induced changes in dopamine neuronal activity.

The effect of glucose on antipsychotic drug-induced changes in the spontaneous activity of rat midbrain dopaminergic neurons was tested with the cells-per-track extracellular electrophysiology method. After daily s.c. treatment with vehicle, haloperidol (0.5 mg/kg), or clozapine (20 mg/kg) for 21 days, rats were anesthetized and recordings performed on one side of the brain. Then, glucose (250 mg/kg, i.v.) was administered and recordings were made from the contralateral midbrain. Glucose significantly reduced the number of spontaneously active A9 and A10 dopaminergic cells per track in control rats, but significantly attenuated the chronic haloperidol- and clozapine-induced reductions in dopaminergic cells per track. These results suggest that caloric intake may influence antipsychotic drug-induced changes in the population activity of midbrain dopaminergic neurons.

Effects of forebrain microinjection of cholecystokinin on dopamine cell firing rate.

In anesthetized rats, midbrain dopamine (DA) neuronal firing rate was differentially sensitive to focal brain microinjection of cholecystokinin peptides (CCK-4 and CCK-8) and N-methyl-D-aspartate (NMDA) into nucleus accumbens, amygdala and prefrontal cortex. Whereas changes in DA neuronal firing rate were frequently observed in response to intra-amygdalar microinjection of CCK peptides, NMDA was most effective in eliciting changes in DA neuronal activity following intra-accumbal microinjection. Thus, stimulation of amygdalar CCK receptors and accumbal excitatory amino acid receptors may participate in the afferent regulation of midbrain DA neuronal function.

Overflow of dopamine and cholecystokinin in rat nucleus accumbens in response to acute drug administration.

Cholecystokinin octapeptide (CCK-8) coexists with dopamine (DA) in rat mesencephalic neurons that project to the nucleus accumbens. To obtain indices of corelease, microdialysis probes were placed in the posterior nucleus accumbens of anesthetized rats, which were then injected acutely (s.c.) with drugs that exert known effects on DA neuronal function. Microdialysis samples were assayed for DA and CCK-8 to determine the differential overflow of these cotransmitters in response to drug treatment. Haloperidol (0.5 mg/kg), d-amphetamine (1 mg/kg), and TCP (5 mg/kg) preferentially increased DA overflow, whereas morphine (5 mg/kg) elicited marked increases in the overflow of both DA and CCK-8. These results suggest that the release of accumbal DA and CCK-8 can be differentially regulated by drug treatment.

Effect of steam-flaked sorghum grain density on performance, mill production rate, and subacute acidosis in feedlot steers.

Two trials were conducted to determine the effects of steam-flaked sorghum grain bulk density on animal performance, cost of production, and propensity to induce ruminal acidosis in feedlot steers. In Trial 1, 336 yearling steers (343 kg; SEM = .346) were fed diets for 125 d that contained sorghum grain (82.5%, DM basis) flaked to .283 (L), .322 (M), or .361 (H) kg/L (i.e., 22, 25, and 28 lb/bu). Steers fed L consumed 3.2% less DM than those fed H (linear, P < .05), resulting in 6.9% lower ADG (linear, P = .02) and 3.6% lower gain efficiency (linear, P < .15). Sorghum grain flaked to M and L had 16 and 46% greater starch gelatinization than H (measured using differential scanning calorimetry; linear, P = .002). Dressing percentage increased linearly (P < .05) with increasing flake density, but no other carcass measurements were affected by treatment. Increasing flake density increased mill production rate linearly (P < .01), resulting in the lowest energy usage per unit of flaked grain for the H treatment. Trial 2 was an acidosis challenge study that incorporated six ruminally cannulated steers (422 kg; SEM = .129) into a replicated 3 x 3 Latin square experiment. Reducing flake density resulted in linear reductions in ruminal pH following intake challenge at 3, 33, and 36 h after the d-12 challenge (P < .05). There was a linear increase in the area between the pH vs time curve and a line at pH 5.5 (P < .01) and 5.0 (P = .09) with decreasing flake density (28.0, 25.2, and 18.2 pH-hours below 5.5 and 9.6, 7.3, and 3.9 pH-hours below 5.0 for L, M, and H, respectively). Cattle consuming L also tended to have higher VFA concentrations (mM) at 36 h after challenge (P = .12). There was no significant treatment effect on ruminal lactate. Flaking sorghum grain to .283 and .322 kg/L resulted in reduced intake and poorer animal performance compared with .361 kg/L (58.7% starch gelatinization), higher susceptibility to subacute acidosis, and higher costs of production.

Effects of electrical stimulation of the central nucleus of the amygdala on the in vivo electrophysiological activity of rat nigral dopaminergic neurons.

The central nucleus of the amygdala (CeA) receives a dopaminergic (DA) innervation from the midbrain. Among its many efferent projections, the CeA innervates the substantia nigra. The possibility that the CeA influences the activity of nigral DA neurons was evaluated. The effects of electrical stimulation of the CeA on the firing rate and pattern of nigral DA neurons were investigated in anesthetized rats. Poststimulus time histograms revealed that nigral DA cells were either inhibited (N = 15), excited (N = 13), or unresponsive (N = 17) to CeA stimulation (250 stimuli at 0.5 Hz). The mean (+/- SEM) latency to inhibition (24 +/- 9 msec) was significantly shorter than that for excitation (65 +/- 10 msec); the duration of inhibition (200 +/- 29 msec) was also significantly greater than the duration of excitation (86 +/- 11 msec) (P < 0.01 for both). DA cells that were excited had basal firing rates significantly lower than those of the inhibited or unresponsive cells (P < 0.05). Preliminary data suggest that DA cell burst-firing increases or decreases, respectively, in association with stimulation-evoked increases or decreases in firing rate. The relatively long latencies for stimulation-evoked responses suggest that CeA projection neurons indirectly affect nigral DA neurons via polysynaptic pathways. These results demonstrate that the CeA has the ability to influence the activity of nigral DA neurons, consistent with the putative role of the CeA as an interface between the limbic and extrapyramidal systems. Given the crucial role of the amygdala in anxiety states, these findings suggest that DA cell function may also be affected in such disorders.

Repeated stimulation of dopamine D2-like receptors: reduced responsiveness of nigrostriatal and mesoaccumbens dopamine neurons to quinpirole.

Extracellular recording techniques were used to study antidromically activated nigrostriatal (NSDA) and mesoaccumbens (MADA) dopamine neurons in chloral hydrate-anesthetized rats. Repeated 14-day i.p. treatment with the dopamine D2-like receptor agonists, quinpirole (2 mg/kg/day) or EMD 23448 (2.6 mg/kg/day), resulted in a significant decrease in the average potency and efficacy of i.v. quinpirole (cumulative doses administered on day 15) to inhibit the spontaneous activity of NSDA neurons relative to vehicle controls. Repeated 14-day quinpirole treatment caused a significantly greater decrease in the sensitivity of MADA neurons to i.v. quinpirole challenges than NSDA neurons. When the effects on NSDA neurons were examined after a shorter treatment period, the decrease in the average potency and efficacy of i.v. quinpirole appeared to occur after only 2 days of i.p. quinpirole treatment (2 mg/kg/day). Iontophoretic studies, however, indicated that the average dopamine sensitivity of somatodendritic dopamine autoreceptors on MADA neurons, but not NSDA neurons, was significantly lower relative to controls after 14-day quinpirole treatment (2 mg/kg/day). These results suggest that this quinpirole treatment regimen can differentially affect the average sensitivity of somatodendritic dopamine autoreceptors on MADA and NSDA neurons. The somatodendritic autoreceptors on MADA neurons appear to be more sensitive to the effects of repeated 14-day quinpirole treatment than those on NSDA neurons.

Effects of administration of cholecystokinin into the VTA on DA overflow in nucleus accumbens and amygdala of freely moving rats.

The carboxyterminal octapeptide of cholecystokinin (CCK-8) coexists with dopamine (DA) in mesolimbic neurons of the ventral tegmental area (VTA). In the present study, in vivo microdialysis in freely moving rats was used to assess the relative effects of sulfated CCK-8 (CCK-8S), unsulfated CCK-8 (CCK-8US) and CCK tetrapeptide (CCK-4), focally injected into the VTA, on DA overflow in two mesolimbic DA/CCK-8S terminal regions, the nucleus accumbens and the amygdala. Consistent with electrophysiological findings, microinjection of CCK-8S, but not CCK-8US or CCK-4, elicited increases in DA overflow in both terminal regions. In the absence of anatomical evidence of CCK-containing fibers in the VTA region, it seems reasonable to conclude that the modulation of terminal DA overflow by CCK-8S through actions at the somatodendritic region represents a form of autoregulation of these cells. Whereas CCK-8US and CCK-4 are preferential CCK-B receptor agonists, CCK-8S binds non-selectively to CCK-A and CCK-B receptors. Thus, these results implicate CCK-A receptors in the stimulatory effects of CCK-8S on VTA DA neurons.

Electrophysiological effects of cholecystokinin on neurons in rat substantia nigra pars reticulata.

The electrophysiological effects of carboxyterminal fragments of cholecystokinin (CCK) on neurons in the dorsal substantia nigra pars reticulata (SNr) were evaluated in anesthetized rats. Microiontophoretic administration of sulfated CCK octapeptide (CCK-8S, agonist for CCK-A and CCK-B receptors) and the selective CCK-B receptor agonists, CCK-4 and unsulfated CCK-8, inhibited the firing rates of a subpopulation of SNr neurons. This effect appears to be mediated by CCK-B receptors.

Influence of excitatory amino acid receptor subtypes on the electrophysiological activity of dopaminergic and nondopaminergic neurons in rat substantia nigra.

In vivo electrophysiological recording methods were used to evaluate the effects of selective and nonselective agonists for excitatory amino acid (EAA) receptor subtypes on the activity of dopaminergic (DA) and nondopaminergic (non-DA) neurons in the substantia nigra of chloral hydrate-anesthetized rats. Microiontophoretic administration of (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA), 2-carboxy-4-(1-methyl-ethenyl)-3-pyrrolidinacetate (kainate), N-methyl-D-aspartate (NMDA) and glutamate excited neurons with an apparent rank order of potency of AMPA = kainate = glutamate > NMDA on DA neurons, and AMPA = kainate > glutamate = NMDA on non-DA neurons. These agonists also changed the firing pattern of DA neurons, which displayed an increase in burst-firing and a reduction in the regularity of the firing pattern. Regularity of firing was indexed by the variation coefficient of a sample of interspike intervals. The apparent potencies of the four agonists to increase burst-firing and variation coefficient were similar to their potencies to increase neuronal firing rate. Blockade of NMDA receptor function by coiontophoresis of 5R,10S-(+/-)-5-methyl-10,11-dihydro-5H-dibenzo[and]cyclohepten-5,1 0-imine hydrogen maleate (MK-801), a selective noncompetitive NMDA antagonist, did not alter kainate-induced changes in firing rate and firing pattern, which indicated that kainate-induced increases in burst-firing were not dependent on concomitant NMDA receptor activation by endogenous excitatory amino acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Further characterization of the effects of BMY 14802 on dopamine neuronal activity.

Further evaluation of the effects of BMY 14802 on dopamine (DA) neuronal activity in the rat substantia nigra pars compacta (A9) was conducted with single-unit recording and microiontophoresis in anesthetized rats. Microiontophoretic administration of BMY 14802 (sigma, serotonin (5-HT)-1A and alpha-1 adrenoceptor ligand) had no effect on DA neurons. Microiontophoretic administration of (+)-3-PPP (weak D2 agonist with high affinity for sigma receptors) and quinpirole (D2/D3 agonist) inhibited A9 DA neuronal activity. Co-iontophoresis or i.v. pretreatment with BMY 14802 had no effect on the current-response curves for the effects of microiontophoretic (+)-3-PPP or quinpirole on A9 DA neurons. Co-iontophoretic administration of (-)-sulpiride, a selective D2 antagonist, blocked the inhibitory effects of microiontophoretic (+)-3-PPP. The effects of BMY 14802 (0.25-8 mg/kg, i.v.) on DA neurons (increased firing rate, increased burst-firing, reduced regularity of firing pattern) were not altered by acute brain hemitransection, but were blocked by pretreatment with NAN-190, an antagonist of 5-HT-1A and alpha-1 receptors. The alpha-1 receptor antagonist, prazosin, did not block these effects of BMY 14802. In conclusion, the effects of BMY 14802 on DA neuronal firing rate and firing pattern are indirect, perhaps due in part to the occupation of 5-HT-1A receptors.

Repeated administration of Sigma ligands alters the population activity of rat midbrain dopaminergic neurons.

Acute and repeated administration of antipsychotic drugs produce distinctive profiles of electrophysiological effects on the population activity of midbrain dopaminergic (DA) neurons which correlate with their clinical effects. Sigma receptors have been hypothesized to be involved in psychosis and in the efficacy of antipsychotic drugs, but little is known about the effects of repeated treatment with sigma ligands on the activity of midbrain DA neuronal populations. In the present study, the cells-per-track cell-sampling method was used to evaluate the effects of 3 sigma ligands on the numbers of spontaneously active A9 and A10 DA neurons in chloral hydrate-anesthetized rats. One-hour pretreatment with either (+)-pentazocine (10 mg/kg, i.p.), DTG (2 mg/kg, i.p.), or JO 1784 (1 or 10 mg/kg, s.c.) did not alter the number of spontaneously active DA neurons encountered per electrode track. Repeated treatment (21 daily injections) with (+)-pentazocine (1 or 10 mg/kg) or DTG (0.2 or 2 mg/kg) increased the number of A10 DA cells per track; JO 1784 (10 mg/kg but not 1 mg/kg) moderately decreased the number of active A9 DA cells and increased the firing rate of A10 DA neurons. The effect of JO 1784 on A9 DA neurons was not due to depolarization inactivation. The effects of all 3 sigma ligands differ from those of antipsychotic drugs, all of which inactivate A10 DA neurons after repeated treatment. Clinical studies are necessary to determine if selective sigma ligands will provide a novel alternative to DA antagonists in the treatment of psychosis.

Repeated amphetamine administration: role of forebrain in reduced responsiveness of nigrostriatal dopamine neurons to dopamine agonists.

The effects of repeated amphetamine treatment on single antidromically identified nigrostriatal dopamine-containing (NSDA) neurons were evaluated in rats. The inhibitory potency and efficacy of dopamine (DA) agonists on NSDA neuron spontaneous discharge rate were examined after amphetamine treatment. Repeated amphetamine treatment (14 days, 1 or 6 mg/kg/day i.p.) dose-dependently decreased the sensitivity of NSDA neurons to the inhibitory effects of the i.v. administered quinpirole. The amphetamine-induced alteration in sensitivity to apomorphine and quinpirole was abolished by acute hemitransection of the forebrain/midbrain connections. No change in the responsiveness or sensitivity of NSDA neurons to the inhibitory effects of iontophoretically applied DA was detected after amphetamine treatment for 14 days (4 or 6 mg/kg/day) or 28 days (6 mg/kg/day). These results suggest that these amphetamine regimens alter the sensitivity/responsiveness of forebrain DA receptors but not NSDA cell somatodendritic DA autoreceptors in a dose-dependent manner.

Ascending afferent regulation of rat midbrain dopamine neurons.

Standard, extracellular single-unit recording techniques were used to examine the electrophysiological and pharmacological responsiveness of midbrain dopamine (DA) neurons to selected, ascending afferent inputs. Sciatic nerve stimulation-induced inhibition of nigrostriatal DA (NSDA) neurons was blocked by both PCPA (5-HT synthesis inhibitor) and 5,7-DHT (5-HT neurotoxin), suggesting mediation by a serotonergic (5-HT) system. Direct stimulation of the dorsal raphe (which utilizes 5-HT as a neurotransmitter and inhibits slowly firing NSDA neurons) inhibited all mesoaccumbens DA (MADA) neurons tested. Paradoxically, DPAT, a 5-HT1A agonist which inhibits 5-HT cell firing, enhanced sciatic nerve stimulation-induced inhibition of NSDA neurons. MADA neurons were not inhibited by sciatic nerve stimulation and, therefore, could not be tested in this paradigm. In contrast to the dorsal raphe, electrical stimulation of the pedunculopontine tegmental nucleus preferentially excited slowly firing NSDA and MADA neurons. Thus, both excitatory and inhibitory ascending afferents influence the activity of midbrain DA neurons, and intact 5-HT systems are necessary for sciatic nerve stimulation to alter DA cell activity. However, the role that 5-HT plays in mediating peripheral sensory input remains unclear.

Effects of phencyclidine, MK-801 and 1,3-di(2-tolyl)guanidine on non-dopaminergic midbrain neurons.

The effects of i.v. administration of the noncompetitive NMDA receptor antagonists, phencyclidine and MK-801, and the sigma receptor ligand, 1,3-di(2-tolyl)guanidine (DTG), on the firing rates of non-dopaminergic mid brain neurons were evaluated in chloral hydrate-anesthetized rats. Phencyclidine and MK-801 inhibited the activity of putative gamma-aminobutyric acid (GABA)-containing interneurons identified by their response to foot-pinch. DTG did not significantly alter neuronal activity. These results suggest that the reported excitatory effects of non-competitive NMDA receptor antagonists on dopamine neuronal activity are due, in part, to disinhibition secondary to the inhibition of interneuron activity.

Electrophysiological effects of MK-801 on rat nigrostriatal and mesoaccumbal dopaminergic neurons.

The electrophysiological effects of the non-competitive N-methyl-D-aspartate (NMDA) antagonist (+)-MK-801 (MK-801) on nigrostriatal and mesoaccumbal dopaminergic (DA) neurons were evaluated in chloral hydrate-anesthetized rats. MK-801 (0.05-3.2 mg/kg, i.v.) stimulated the firing rates of 14 (74%) of 19 nigrostriatal DA (NSDA) neurons and all 16 mesoaccumbal DA (MADA) neurons tested. Stimulatory effects of the drug were more prominent on MADA neurons. Interspike interval analysis revealed that MK-801 also regularized DA neuronal firing pattern. Acute brain hemitransection between the midbrain and forebrain attenuated the stimulatory effects of MK-801 on firing rate and blocked the effects on firing pattern. Similar to MK-801, hemitransection itself increased NSDA and MADA cell firing rates and regularized firing pattern. Both i.v. and iontophoretic MK-801 blocked the excitatory effects of iontophoretic NMDA but did not affect excitations caused by the non-NMDA glutamatergic receptor agonists quisqualate and kainate. Iontophoretic MK-801 had no effect alone. These results suggest that the excitatory effects of i.v. MK-801 on DA neuronal activity are not due to direct actions on DA neurons. Glutamatergic projections originating anterior to the hemitransection appear to play a role in the effects of MK-801 on DA neuronal activity.

Acute effects of sigma ligands on the electrophysiological activity of rat nigrostriatal and mesoaccumbal dopaminergic neurons.

The effects of acute i.v. administration of several sigma ligands on the single-unit activity of nigrostriatal and mesoaccumbal dopaminergic (DA) neurons were evaluated in chloral hydrate-anesthetized rats. DTG (1,3-di(o-tolyl)guanidine) did not alter DA neuronal activity at nontoxic doses and JO 1784 [(+)-N-cyclopropylmethyl-N-methyl-1,4-diphenyl-1-ethylbut-3-en-1-+ ++ylamine] was inactive. (+)-Pentazocine was more effective in increasing mesoaccumbal vs. nigrostriatal DA cell firing rates. BMY 14802(alpha-(4-fluorophenyl)-4-(5-fluoro-2-pyrimidinyl)-1-piperazine-but anol) dose-dependently increased DA cell firing rate in both populations. The inhibition of nigrostriatal DA cell firing rate by (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine [(+)-3-PPP] was reversed by (-)-eticlopride and (+)-but not (-)-butaclamol, which supports previous evidence that (+)-3-PPP-induced inhibition is due to the DA agonist properties of the drug. From what is known of the pharmacological properties of these compounds, it is concluded that acute sigma receptor occupation does not markedly alter the firing rate of DA neurons. The dose-response curve for inhibition of nigrostriatal DA neuronal activity by the D2 DA agonist, quinpirole, was shifted to the right tenfold by BMY 14802 pretreatment (8 mg/kg, i.v.) and twofold by (+)-pentazocine (8 mg/kg, i.v.), but was not changed by DTG (2 mg/kg, i.v.). It is concluded that the marked effects of certain sigma ligands on DA cell electrophysiology are likely due to their non-sigma properties.

Effects of supplemental protein percentage and feeding level on intake, ruminal fermentation, and digesta passage in beef steers fed prairie hay.

Twelve ruminally cannulated steers (average initial BW 357 kg) were allotted to four treatments (three steers per treatment) in a replicated 4 x 4 Latin square design with 21-d periods (12 d for adaptation and 9 d for collection) to compare the effects of protein supplements that differed in percentage of CP and feeding level on low-quality forage utilization. Treatments were 1) control (C), ad libitum access to 5.6% CP prairie hay, 2) C +600 g of DM.steer-1.d-1 of a 43% CP supplement based on cottonseed meal (PS), 3) C + 1,200 g of DM.steer-1.d-1 of a 22% CP supplement based on corn grain and cottonseed meal (GS), and 4) C + 600 g of DM.steer-1.d-1 of a 22% CP supplement based on corn grain and cottonseed meal (LS). Ruminal total VFA concentrations were increased 8% (P less than .07) by PS vs GS 1 h after supplementation. Among supplemented steers, ruminal acetate (mol/100 mol) was decreased 1.2 mol/100 (P less than .03) by GS vs PS and LS; however, supplementation did not affect (P greater than .10) acetate proportions compared with C. Neither propionate nor butyrate was affected (P greater than .10) by supplementation, but among supplemented steers, butyrate proportions were 8% greater (P less than .03) for GS than for PS and 5% less (P less than .10) for LS than for the average of GS and PS. Ruminal pH did not differ (P greater than .10) among treatments. Ruminal ammonia concentrations were increased 1.4 to 4.8 mg/100 mL (P less than .07) by supplementation and typically were less for LS than for PS and GS at most sampling times. Prairie hay DMI (average = 16.3 g/kg BW) was not affected (P greater than .10) by supplementation. Fluid dilution rate was 8% faster (P less than .01) when steers were supplemented than when they were not fed supplement, and fluid dilution rate was increased 4% (P less than .04) by GS compared with PS. Particulate digesta passage rate was not affected (P greater than .10) by treatment, but total tract retention time was decreased (P less than .01) 10% by supplementation. Extent and rate of prairie hay NDF digestion in situ were not greatly affected by supplementation, but in situ disappearance of supplement N was 6 to 10 percentage units less (P less than .06) for GS than for PS and 2 to 6 percentage units less for LS than for the average of PS and GS supplements.(ABSTRACT TRUNCATED AT 400 WORDS)