Epithelial Blood Flow Research Articles

Ergot alkaloids from endophyte-infected tall fescue decrease reticuloruminal epithelial blood flow and volatile fatty acid absorption from the washed reticulorumen1,2

An experiment was conducted to determine if ergot alkaloids affect blood flow to the absorptive surface of the rumen. Steers (n=8) were pair-fed alfalfa cubes and received ground endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum; E+) seed (0.015 mg ergovaline·kg BW(-1)·d(-1)) or endophyte-free tall fescue (E-) seed via the rumen cannula 2x daily for 7 d at thermoneutral (TN; 22°C) and heat stress (HS; 32°C) conditions. On d 8, the rumen was emptied and rinsed. A buffer containing VFA was incubated in the following sequence: control (CON), 15 μg ergovaline·kg BW(-1) (1×EXT) from a tall fescue seed extract, and 45 μg ergovaline·kg BW(-1) (3×EXT). For each buffer treatment there were two 30-min incubations: a 30-min incubation of a treatment buffer with no sampling followed by an incubation of an identical sampling buffer with the addition of Cr-EDTA and deuterium oxide (D2O). Epithelial blood flow was calculated as ruminal clearance of D2O corrected for influx of physiological water and liquid outflow. Feed intake decreased with dosing E+ seed at HS but not at thermoneutral conditions (TN; P<0.02). Dosing E+ seed decreased serum prolactin (P<0.005) at TN. At HS, prolactin decreased in both groups over the 8-d experiment (P<0.0001), but there was no difference in E+ and E- steers (P=0.33). There was a seed treatment×buffer treatment interaction at TN (P=0.038), indicating that E+ seed treatment decreased reticuloruminal epithelial blood flow at TN during the CON incubation, but the two groups of steers were not different during 1×EXT and 3×EXT (P>0.05). Inclusion of the extract in the buffer caused at least a 50% reduction in epithelial blood flow at TN (P=0.004), but there was no difference between 1×EXT and 3×EXT. There was a seed × buffer treatment interaction at HS (P=0.005), indicating that the reduction of blood flow induced by incubating the extract was larger for steers receiving E- seed than E+ seed. Volatile fatty acid flux was reduced during the 1×EXT and 3×EXT treatments (P<0.01). An additional experiment was conducted to determine the effect of time on blood flow and VFA flux because buffer sequence could not be randomized. Time either increased (P=0.05) or did not affect blood flow (P=0.18) or VFA flux (P>0.80), indicating that observed differences are due to the presence of ergot alkaloids in the rumen. A decrease in VFA absorption could contribute to the signs of fescue toxicosis including depressed growth and performance.

A model of ruminal volatile fatty acid absorption kinetics and rumen epithelial blood flow in lactating Holstein cows

Ruminal absorption of volatile fatty acids (VFA) is quantitatively the most important nutrient flux in cattle. Historically, VFA absorption models have been derived primarily from ruminal variables such as chemical composition of the fluid, volume, and pH. Recently, a mechanistic model incorporated the control of VFA absorption from epithelial surface area of the reticulorumen. In the present study, we hypothesized that ruminal absorption of VFA was controlled through epithelial permeability to VFA and rumen epithelial capillary blood flow. The objective of the study was to construct a model of VFA exchange across the rumen wall that incorporates epithelial blood flow as a driving force for ruminal VFA removal. The bidirectional fluxes between the ruminal and epithelial pool of VFA were assumed mass action driven, given that passive diffusion of nonionized VFA is the dominant transmembrane VFA flux. Parameter estimates were derived by fitting the model to observed data. The model provided reliable unbiased estimates of ruminal VFA absorption and rumen epithelial blood flow. Blood flow was modeled using an equation that considered the effect of butyrate and dietary crude protein intake per kilogram of body weight. The rate constants related to the flux from ruminal fluid to epithelium were in the order isobutyrate < acetate < propionate < butyrate (0.32±0.02, 0.72±0.2, 0.91±0.06, and 0.97±0.02 /h, respectively). The rate constants for fluxes of isobutyrate, acetate, propionate, and butyrate from the rumen epithelium to the ruminal fluid, relative to the pool size of the epithelium, were 4.78, 10.6, 13.4, and 14.3 /h, respectively. Ruminal concentrations of acetate, propionate, butyrate, and isobutyrate were predicted with root mean square prediction errors as percentage of the observed means (RMSPE) of 5.86, 5.75, 11.3, and 4.12, respectively. The epithelial blood flow was predicted with 26.3% RMSPE. Sensitivity analyses indicated that when ruminal butyrate concentration increased from 4.0 to 37.4mmol/L, blood flow of the epithelium increased 47% and the ruminal disappearance rate of propionate increased 11%. The concentration gradient of propionate between ruminal fluid and epithelium was no more than 3:1 and increased with increasing blood flow. In conclusion, a dynamic model based on rumen epithelial blood flow and bidirectional fluxes of VFA between ruminal fluid and epithelium gave unbiased predictions with low residual error of ruminal VFA absorption under washed rumen conditions. The model indicates that the effect of varying epithelial blood flow on the control of ruminal VFA absorption is related to the concentration gradient of individual VFA between ruminal fluid and epithelial blood. Epithelial blood flow may be an important determinant of ruminal absorption of VFA, a result that has not been evaluated on independent data.

Effects of ruminal ammonia and butyrate concentrations on reticuloruminal epithelial blood flow and volatile fatty acid absorption kinetics under washed reticulorumen conditions in lactating dairy cows

The effect of reticuloruminal epithelial blood flow on the absorption of propionate as a volatile fatty acid (VFA) marker in 8 lactating Holstein cows was studied under washed rumen conditions. The cows were surgically prepared with ruminal cannulas and permanent catheters in an artery and mesenteric, right ruminal, and hepatic portal veins. The experiment was designed with 2 groups of cows: 4 cows adapted to high crude protein (CP) and 4 to low CP. All cows were subjected to 3 buffers: butyric, ammonia, and control in a randomized replicated 3 × 3 incomplete Latin square design. The buffers (30kg) were maintained in a temporarily emptied and washed rumen for 40min. The initial concentration of VFA was 84.2mmol/L. Butyrate was increased from 4 to 36mmol/L in butyric buffer by replacement of acetate, and ammonia (NH3) was increased from 2.5 to 22.5mmol/L in ammonia buffer by replacement of NaCl. Increasing amounts of deuterium oxide (D2O) were added to the buffers as the order of buffer sequence increased (6, 12, and 18g of D2O). Ruminal clearance of D2O was used to estimate epithelial blood flow. To increase accuracy of the epithelial blood flow estimates, data of ruminal liquid marker (Cr-EDTA), and initial and final buffer volumes were fitted to a dynamic simulation model. The model was used to estimate ruminal liquid passages, residual liquid, and water influx (saliva and epithelia water) for each combination of cow and buffer (n=24). Epithelial blood flow increased 49±11% for butyric buffer compared with control. The ruminal disappearance of propionate (marker VFA) was affected by buffer and followed the same pattern as for epithelial blood flow. The correlation between ruminal disappearance of propionate and epithelial blood flow (r=0.56) indicates that the removal of propionate can be limited by epithelial blood flow. The ruminal disappearance of propionate increased 30±12% for the butyric compared with ammonia buffer and 12.5±8% when compared with control. The net portal flux of propionate increased 32±6% in butyric compared with control. In conclusion, rumen epithelial blood flow is positively correlated with ruminal disappearance of propionate and affects the kinetics of ruminal VFA absorption.

Characterization of cytochrome P-450-dependent arachidonic acid metabolism in rabbit intestine

We characterized cytochrome P-450-dependent arachidonate (P-450-AA) metabolism throughout the intestinal tract, since some metabolites derived via this pathway modify epithelial ion transport and regional blood flow. Microsomes (0.3 mg/ml) were prepared from each region of the intestines of anesthetized New Zealand White male rabbits and incubated with [14C]AA (7 microM) for 30 min at 37 degrees C. In the presence of NADPH (1 mM), ileal microsomes exhibited the greatest P-450-AA metabolism, whereas duodenal microsomes exhibited little or no activity. For jejunal, ileal, and cecal microsomes, AA metabolism was reduced in the absence of NADPH and by boiling microsomes, was unaffected by indomethacin (10 microM) and BW-755C (50 microM), but was significantly attenuated by the P-450 enzyme inhibitors, 7-ethoxyresorufin (1 microM) and SKF-525A (100 microM). However, colonic (ascending, transverse, and descending) microsomal activity was inhibited by both P-450 and lipoxygenase inhibitors. Analysis of ileal AA metabolites by high-pressure liquid chromatography and negative ion chemical ionization gas chromatography-mass spectrometry revealed products corresponding to monohydroxyeicosatetraenoic acids (HETEs). Semiquantitative analysis showed that 20-, 19-, 18-, 17-, and 16-HETEs were present in a ratio of 6.2:3.3:0.3:0.1:0.1, respectively. Furthermore, ileal P-450-HETEs dilated the isolated perfused mesenteric bed, as did 20-HETE, the predominant ileal AA metabolite. Because 20-HETE was also shown to affect epithelial ion transport, we suggest that P-450-AA metabolites may make important contributions to intestinal function.