Comparison of pH and bacterial communities in the rumen and reticulum during fattening of Japanese Black beef cattle.

We used castrated and fistulated Japanese Black beef cattle (n = 9) to measure the pH and bacterial communities in the rumen liquid, rumen solid, and reticulum liquid during early, middle, and late fattening stages (10-14, 15-22, and 23-30 months of age, respectively). The pH was measured in the rumen and reticulum during the last 13 days of each fattening stage and was significantly lower in the rumen at the early and middle fattening stage and in the reticulum during the late stage. Sequencing analysis indicated similar bacterial compositions in the rumen and reticulum liquid fractions and stability of bacterial diversity in the rumen and reticulum liquid fractions and rumen solid fraction. By contrast, major operational taxonomic units (OTUs), such as Ruminococcus bromii strain ATCC 27255 (OTU1, OTU10, and OTU15), were differently correlated to the fermentation parameters among the rumen and reticulum liquid fractions. Therefore, the long-term feeding of Japanese Black beef cattle with a high-concentrate diet might reverse the trend of pH in the rumen and reticulum during the late fattening stage, and the bacterial communities adapted to changes in fermentation by preserving their diversity throughout fattening.

Modeling reticular and ventral ruminal pH of lactating dairy cows using ingestion and rumination behavior.

The prevention and control of metabolic and digestive diseases is an enormous challenge in dairy farming. Subacute ruminal acidosis (SARA) is assumed to be the most severe feed-related disorder and it impairs both animal health and economic efficiency. Currently, ruminal pH as well as variables derived from the daily pH curve are the main indicators for SARA. The objective of this study was to explain the daily pH course in the ventral rumen and reticulum of dairy cows using ingestion pattern and rumination behavior data gathered by automated data recording systems. The data of 13 ruminally fistulated lactating cows were collected at the experimental station of the Friedrich-Loeffler-Institut (Brunswick, Germany). The data included continuous pH measurements, which were recorded simultaneously in the reticulum by pH-measuring boluses and in the ventral rumen by a separate data logger. In addition, rumination behavior was measured using jaw movement sensors, and feed and water intakes were recorded by transponder-assisted systems. Milk yield and body weight were determined during and after each milking, respectively. For statistical evaluation, the data were analyzed using time-series modeling with multiple linear mixed regressions. Before applying the developed mathematical statistical modeling, we performed a plausibility assessment to ensure data quality. The major part of the mathematical statistical modeling consisted of data preparation, where all variables were transformed into a uniform 1-min resolution. Signal transformations were used to model individual feed and water intakes as well as rumination behavior events over time. Our results indicated that diurnal pH curves of both the reticulum and ventral rumen could be predicted by the transformed feed and water intake rates. Rumination events were associated with a marginal temporal increase in pH. We observed that the pH of the ventral rumen was delayed by approximately 37 min compared with that of the reticulum, which was therefore considered in the modeling. With the models developed in this study, 67.0% of the variance of the reticular pH curves and 37.8% of the variance of the ruminal pH curves could be explained by fixed effects. We deduced that the diurnal pH course is, to a large extent, associated with the animal's individual feed intake and rumination behavior.

Prenatal histomorphological development of the reticulum in fallow deer (Dama dama).

The histomorphological changes occurring in the Dama dama reticulum during prenatal development have been investigated. Twenty-five Dama dama embryos were used, from the first stages of prenatal life until birth. Differentiation of the reticulum was observed at 23% gestation. By 25% gestation the reticular wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Primary reticular crests were visible at 38% gestation. Secondary reticular crests were observed at 61% gestation. Neuroendocrine cells were detected by synaptophysin (SYP) at 35% gestation, in the lamina propria-submucosa, tunica muscularis, and serosa. Epithelial Cytokeratin-18 (CK-18) cells were observed at 35% gestation extended throughout the epithelial layers. The glial cells (vimentin -VIM- and glial fibrillary acidic protein-GFAP-markers) were discerned at 25% and 43% gestation, respectively, in myenteric and submucosal plexuses, lamina propria, muscularis mucosae, tunica muscularis, and perivascular connective tissue. The neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) markers were immunodetected at 75% and 80 gestation, respectively, in the lamina propria-submucosa, muscularis mucosae, tunica muscularis, serosa, and myenteric plexuses. The prenatal development of the fallow deer reticular mucosa evidenced a considerable precocity similar to that previously reported in goat and red deer.

Developing and evaluating threshold-based algorithms to detect drinking behavior in dairy cows using reticulorumen temperature.

In this study, we assessed for the first time the use of a reticuloruminal temperature bolus and a thresholding method to detect drinking events and investigated different factors that can affect drinking behavior. First, we validated the detection of drinking events using 16 cows that received a reticuloruminal bolus. For this, we collected continuous drinking behavior data for 4 d using video recordings and ambient and water temperature for the same 4 d. After all the data were synchronized, we performed 2 threshold algorithms: a general-fixed threshold and a cow-day specific threshold algorithm. In the general-fixed threshold, a positive test was considered if the temperature of any cow fell below a fixed threshold; in the cow-day specific threshold, a positive test was considered when the temperature of specific cows fell below the threshold value deviations around the mean temperature of the cow for that day. The former was evaluated using a threshold varying between 35.7 and 39.5°C, and the latter using the formula µ-n10σ, where µ = mean of the temperature of each cow for one day, n = 1, 2, …, 20, and σ = standard deviation of the temperature of each cow on that day. The performance of the validation of detection using each of the threshold types was computed using different metrics, including overall accuracy, precision, recall (also known as sensitivity), F-score, positive predictive value, negative predictive value, false discovery rate, false omission rate, and Cohen's kappa statistic. The findings of the first study showed that the cow-day specific threshold of n = 10 performed better (true positives = 466; false positives = 167; false negatives = 165; true negatives = 8,416) than using a general-fixed threshold of 38.1°C (true positives = 449; false positives = 181; false negatives = 182; true negatives = 8,402). With the information gained in this first study, we investigated the different factors associated with temperature drop characteristics per cow: number of drops, mean amplitude of the drop, and mean recovery time. For this, we used data from 54 cows collected for almost 1 yr to build a mixed-effect multilevel model that included days in milk, parity, average monthly milk production, and ambient temperature as explanatory variables. Cow characteristics and ambient temperature had significant effects on drinking events. Our results provide a platform for automated monitoring of drinking behavior, which has potential value in prediction of health and welfare in dairy cattle.

Changes in ruminal and reticular pH and bacterial communities in Holstein cattle fed a high-grain diet.

BACKGROUND: Subacute ruminal acidosis (SARA) is characterized by a ruminal pH depression, and microbiota can also be affected by a higher acidity and/or dietary changes. Previous studies have revealed similar patterns in pH reduction in the rumen and reticulum, whereas changes in reticular pH and bacterial community following a high-grain diet are not fully understood. The aim of this study was to investigate the changes in reticular pH and bacterial community structure following a high-grain diet simultaneously with those in the rumen. RESULTS: SARA was diagnosed when ruminal and reticular pH remained under 5.6 for 350 ± 14 and 312 ± 24 min/d, respectively, on the last day of the CON period. During the CON period, lower proportion of acetic acid and higher proportion of butyric acid were observed compared with the HAY period. The proportions of acetic acid and propionic acid were lower and higher, respectively, in the rumen compared with the reticulum. From 454 pyrosequencing analysis, the relative abundance of several genera differed significantly between the two periods and the two locations. During the HAY period, higher relative abundances of Prevotella, Eubacterium, Oscillibacter, and Succiniclasticum and lower relative abundances of Ruminococcus, Clostridium, and Olsenella were identified compared with the CON period. Furthermore, the relative abundance of Eubacterium was lower in the rumen compared with the reticulum. Bacterial diversity indices were significantly different between the HAY and CON periods, being higher in the HAY period. The quantitative real-time PCR showed that the copy numbers of several cellulolytic bacteria (Fibrobacter succinogenes and Ruminococcus albus) were higher during the HAY period. CONCLUSION: A high-grain diet showed similar impacts on the pH, fermentation, and bacterial community structure in the rumen and reticulum. During the CON period, ruminal and reticular pH decreased following the high-grain challenge, and lower bacterial diversity and changes in the bacterial composition, similarity, and bacterial copy numbers were observed due to a higher acidity and dietary changes compared with the HAY period. These changes may influence the fermentative ability of the rumen and reticulum.

Effect of Entodinium caudatum on starch intake and glycogen formation by Eudiplodinium maggii in the rumen and reticulum.

This study aimed to quantify the engulfed starch and reserve α-glucans (glycogen) in the cells of the ciliates Eudiplodinium maggii, as well the α-glucans in defaunated and selectively faunated sheep. The content of starch inside the cell of ciliates varied from 21 to 183mg/g protozoal DM relative to the rumen fauna composition whereas, the glycogen fluctuated between 17 and 126mg/g dry matter (DM) of this ciliate species. Establishment of the population Entodinium caudatum in the rumen of sheep already faunated with E. maggii caused a drop in both types of quantified carbohydrates. The content of α-glucans in the rumen of defaunated sheep varied from 4.4 to 19.9mg/g DM and increased to 7.4-29.9 or 11.8-33.9mg/g DM of rumen contents in the presence of only E. maggii or E. maggii and E. caudatum, respectively. The lowest content of the carbohydrates was always found just before feeding and the highest at 4h thereafter. The α-glucans in the reticulum varied 7.5-40.1, 14.3-76.8 or 21.9-106.1mg/g DM of reticulum content for defaunated, monofaunated or bifaunated sheep, respectively. The results indicated that both ciliate species engulf starch granules and convert the digestion products to the glycogen, diminishing the pool of starch available for amylolytic bacteria.

Determination of Henry's constant, the dissociation constant, and the buffer capacity of the bicarbonate system in ruminal fluid.

Despite the clinical importance of ruminal acidosis, ruminal buffering continues to be poorly understood. In particular, the constants for the dissociation of H2CO3 and the solubility of CO2 (Henry's constant) have never been stringently determined for ruminal fluid. The pH was measured in parallel directly in the rumen and the reticulum in vivo, and in samples obtained via aspiration from 10 fistulated cows on hay- or concentrate-based diets. The equilibrium constants of the bicarbonate system were measured at 38°C both using the Astrup technique and a newly developed method with titration at 2 levels of partial pressure of CO2 (pCO2; 4.75 and 94.98 kPa), yielding mean values of 0.234 ± 0.005 mmol ∙ L(-1) ∙ kPa(-1) and 6.11 ± 0.02 for Henry's constant and the dissociation constant, respectively (n/n = 31/10). Both reticular pH and the pH of samples measured after removal were more alkalic than those measured in vivo in the rumen (by ΔpH = 0.87 ± 0.04 and 0.26 ± 0.04). The amount of acid or base required to shift the pH of ruminal samples to 6.4 or 5.8 (base excess) differed between the 2 feeding groups. Experimental results are compared with the mathematical predictions of an open 2-buffer Henderson-Hasselbalch equilibrium model. Because pCO2 has pronounced effects on ruminal pH and can decrease rapidly in samples removed from the rumen, introduction of a generally accepted protocol for determining the acid-base status of ruminal fluid with standard levels of pCO2 and measurement of base excess in addition to pH should be considered.

Use of dicarboxylic acids and polyphenols to attenuate reticular pH drop and acute phase response in dairy heifers fed a high grain diet.

BACKGROUND: The aim of this study was to determine the ability of two feed additives, a fumarate-malate (FM) and a polyphenol-essential oil mixture (PM), in attenuating the drop of ruminal pH and the metabolic and immune response resulting from an excessively high grain diet. Six heifers were used in a 3 × 3 Latin square experiment and fed a low starch (LS) diet for 14 d, followed by a high starch (HS) diet for 8 d (NDF 33.6%, starch 30.0% DM). In the last 5 days of each period, barley meal was added to decrease rumen pH. During HS feeding all animals were randomly assigned to one of the following three dietary treatments: no supplement/control (CT), a daily dose of 60 g/d of FM, or 100 g/d of PM. Reticular pH was continuously recorded using wireless boluses. On d 21 of each period, rumen fluid was collected by rumenocentesis (1400 h), together with blood (0800 h) and fecal samples (0800, 1400, and 2100 h). RESULTS: The correlation coefficient of pH values obtained using the boluses and rumenocentesis was 0.83. Compared with CT and PM, the FM treatment led to a lower DMI. Nadir pH was lowest during CT (5.40, 5.69, and 5.62 for CT, FM and PM, respectively), confirming the effectiveness of both supplements in reducing the pH drop caused by high grain feeding. This result was confirmed by the highest average time spent daily below 5.6 pH (199, 16 and 18 min/d) and by the highest acetate to propionate ratio of the CT fed heifers. The PM decreased the concentrations of neutrophils (2.9, 3.2, and 2.8 10(9)/L) and acute phase proteins: SAA (37.1, 28.6 and 20.1 µg/mL), LBP (4.1, 3.8, and 2.9 µg/mL), and Hp (675, 695 and 601 µg/mL). Free lipopolysaccharides (LPS) were detected in blood and feces, but their concentrations were not affected by treatments, as the remaining blood variables. CONCLUSIONS: Data suggest that both additives could be useful in attenuating the effects of excessive grain feeding on rumen pH, but the PM supplement was more effective than FM in reducing the inflammatory response compared to CT.

Oxidative stress-induced mitochondrial fragmentation and movement in skeletal muscle myoblasts.

Mitochondria are dynamic organelles, capable of altering their morphology and function. However, the mechanisms governing these changes have not been fully elucidated, particularly in muscle cells. We demonstrated that oxidative stress with H2O2 resulted in a 41% increase in fragmentation of the mitochondrial reticulum in myoblasts within 3 h of exposure, an effect that was preceded by a reduction in membrane potential. Using live cell imaging, we monitored mitochondrial motility and found that oxidative stress resulted in a 30% reduction in the average velocity of mitochondria. This was accompanied by parallel reductions in both organelle fission and fusion. The attenuation in mitochondrial movement was abolished by the addition of N-acetylcysteine. To investigate whether H2O2-induced fragmentation was mediated by dynamin-related protein 1, we incubated cells with mDivi1, an inhibitor of dynamin-related protein 1 translocation to mitochondria. mDivi1 attenuated oxidative stress-induced mitochondrial fragmentation by 27%. Moreover, we demonstrated that exposure to H2O2 upregulated endoplasmic reticulum-unfolded protein response markers before the initiation of mitophagy signaling and the mitochondrial-unfolded protein response. These findings indicate that oxidative stress is a vital signaling mechanism in the regulation of mitochondrial morphology and motility.

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

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.

Kinetics of phosphate absorption in lactating dairy cows after enteral administration of sodium phosphate or calcium phosphate salts.

Hypophosphataemia is frequently encountered in dairy cows during early lactation. Although supplementation of P is generally recommended, controversy exists over the suitability of oral P supplementation in animals with decreased or absent rumen motility. Since the effects of transruminal P absorption and the reticular groove reflex on the absorption kinetics of P are not well understood, it is unclear in how far treatment efficacy of oral P supplementation is affected by decreased rumen motility. Phosphate absorption was studied in six phosphate-depleted dairy cows fitted with rumen cannulas and treated with test solutions containing either NaH2PO4 or CaHPO4 with acetaminophen. Each animal was treated orally, intraruminally and intra-abomasally in randomised order. Absorption kinetics of P were studied and compared with the absorption kinetics of acetaminophen, a marker substance only absorbed from the small intestine. Intra-abomasal treatment with NaH2PO4 resulted in the most rapid and highest peaks in plasma inorganic P (Pi) concentration. Oral and intraruminal administration of NaH2PO4 resulted in similar increases in plasma Pi concentration from 4 to 7 h in both groups. Treatment with NaH2PO4 caused more pronounced peaks in plasma Pi concentration compared with CaHPO4. Neither transruminal P absorption nor the reticular groove reflex affected P absorption kinetics as determined by comparing plasma concentration­time curves of P and acetaminophen after administration of 1M-phosphate salt solutions. It is concluded that oral treatment with NaH2PO4 but not CaHPO4 is effective in supplementing P in hypophosphataemic cows with adequate rumen motility. Decreased rumen motility is likely to hamper the efficacy of oral phosphate treatment.

Simultaneous estimation of the pH of rumen and reticulum fluids of cows using a radio-transmission pH-measurement system.

Circadian pH changes in the fluid of the rumen (bottom and middle) and reticulum were assessed simultaneously using wireless and wired radio-transmission pH-measurement systems in cows fed a control diet (C diet) or rumen-acidosis-inducing diet (RAI diet). The pH in the three sites decreased following the morning and evening feedings. In cows fed the C diet, the bottom-rumen and reticular pH reverted to the basal level by the next morning, while the middle-rumen pH did not recover completely, suggesting that active fermentation occurred in the middle of the rumen. The mean pH at 1 hr intervals was higher in the reticulum than at the bottom and in the middle of the rumen. The relatively stable reticular pH may result from dilution due to salivation. In cows fed the RAI diet, the bottom-rumen pH fell to approximately 5.2 after the evening feeding, but returned to the basal level by the next morning. In contrast, the middle-rumen pH did not return to the basal level (6.5) within 24 hr, presumably owing to continuous, vigorous fermentation. There were positive correlations between the pH at the bottom and in the middle of the rumen and at the bottom of the rumen and in the reticulum. These findings indicate that our radio-transmission pH-measurement system may be suitable tool for simultaneous measurement of pH in the rumen and reticulum fluid.

The use of receiver operating characteristic (ROC) analysis to assess the diagnostic value of serum amyloid A, haptoglobin and fibrinogen in traumatic reticuloperitonitis in cattle.

Acute phase proteins (APPs) such as serum amyloid A (SAA), haptoglobin (Hp), and plasma fibrinogen (Fb) are diagnostic and prognostic biomarkers. This study determined the pattern of SAA in traumatic reticuloperitonitis (TRP) in cows, and compared the findings with Hp, and plasma Fb concentrations in the differential diagnosis of TRP using receiver operating characteristic (ROC) analysis. Blood samples were collected from 89 cows: 53 cases of TRP, 16 animals with other internal disorders and 20 healthy controls. SAA and Hp had a similar pattern and magnitude of changes in TRP. Using the ROC method, the optimal cut-off for the diagnosis of TRP was 68 microg/mL for SAA and 0.74 g/L for Hp, with 100% sensitivity and 86.1% specificity; either of these parameters may therefore be used as a sensitive and relatively specific tool for the differential diagnosis of TRP in cattle. The diagnostic accuracy of plasma Fb was significantly lower than either SAA or Hp (P<0.01). The combined use of these APPs (whilst aware of their conditional interdependence) may not provide any additional diagnostic information than can be obtained using either SAA or Hp alone.

[Dynamics of rumenoenteral transport of progesterone in sheep in chronic experiment].

In chronic experiments on pregnant and lactating sheeps with large fistula of rumen the dynamics of progesterone transport in a cavity of temporarily isolated reticulo-rumen depending on a physiological condition of animals, composition of solution filling a cavity of organ, and time of incubation, were studied. On three animals, 24 experiments were carried out. It was established that, during pregnancy, in a cavity of reticulo-rumen more progesterone was transported than during lactation (p < 0.05). Addition of lignin and cellulose to the salt solution increased the progesterone transport in the cavity of reticulo-rumen (p < 0.001). During three-hour incubation of salt solution in the cavity of reticulo-rumen, an increase of progesterone transport was observed in cavity of the organ (p < 0.001). The data obtained confirm the assumption that forestomach of ruminants participates in removing of progesterone from internal environment to enteral one and that carbohydrate polymers facilitate this process.

Splanchnic metabolism of volatile fatty acids absorbed from the washed reticulorumen of steers.

Six steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, as well as in the right ruminal vein were used to study metabolism of VFA absorbed from buffers in the emptied and washed reticulorumen. [2-(13)C]Acetate was infused into a jugular vein to study portal-drained visceral (PDV) uptake of arterial acetate, hepatic unidirectional uptake of acetate, and whole-body irreversible loss rate (ILR). Isobutyrate was infused into the right ruminal vein to calibrate VFA fluxes measured in the portal vein. On sampling days, the rumen was emptied and incubated in sequence with a 0-buffer (bicarbonate buffer without VFA), a VFA-buffer plus continuous intraruminal infusion of VFA, and finally another 0-buffer. Ruminal VFA absorption was determined as VFA uptake from the VFA-buffer and metabolic effects determined as the difference between metabolite fluxes with VFA-buffer and 0-buffers. Steady absorption rates of VFA were maintained during VFA-buffer incubations (4 h; 592+/-16, 257+/-5, 127+/-2, 17+/-<1, 20+/-<1 mmol/h, respectively, of acetate, propionate, butyrate, isovalerate, and valerate). The portal flux of acetate corrected for PDV uptake of arterial acetate accounted for 105+/-3% of the acetate absorption from the rumen, and the net portal flux of propionate accounted for 91+/-2% of propionate absorption. Considerably less butyrate (27+/-3%) and valerate (30+/-3%) could be accounted for in the portal vein. The sum of portal VFA and 3-hydroxybutyrate as well as lactate represented 99+/-3% of total VFA acetyl units and 103+/-2% of VFA propionyl units. Estimates are maximum because no accounting was made for lactate derived from glycolysis in the PDV. The net splanchnic flux of VFA, lactate, 3-hydroxybutyrate, and glucose accounted for 64+/-2% of VFA acetyl units and 34+/-5% of VFA propionyl units. Results indicate that there is a low "first-pass" uptake of acetate and propionate in the ruminal epithelium of cattle, whereas butyrate and valerate are extensively metabolized, though seemingly not oxidized to carbon dioxide in the epithelium but repackaged into acetate, 3-hydroxybutyrate, and perhaps other metabolites. When PDV "second-pass" uptake of arterial nutrients is accounted for, PDV fluxes of VFA, lactate, and 3-hydroxybutyrate represent VFA production in the gastrointestinal tract and thereby VFA availability to the ruminant animal.

The absorption of calcium ions from the ovine reticulo-rumen.

Net Ca2+ and Mg2+ absorption rates were measured in vivo from buffer solutions placed in the washed reticulo-rumen, isolated in situ in 30 conscious, trained sheep. An increase in concentration of short chain fatty acids (SCFA) in the buffer, over the range 0-50 mM, was shown to stimulate the net rates of absorption of Ca2+ and Mg2+ ions from the rumen. Similarly, the results of in vitro experiments, carried out with ovine rumen epithelium mounted in short-circuited Ussing chambers, showed that the absence of SCFA from the chamber fluid resulted in a reduction in Jnet Ca2+ caused by reduced flux of Ca2+ ions in the mucosal to serosal direction (Jms Ca2+). The addition of 1 mM acetazolamide, an inhibitor of carbonic anhydrase, to the ruminal buffer used in the in vivo experiments led to significant reductions in the net absorption rates of Ca2+ and Mg2+ ions in the presence of SCFA (50 mmol x l(-1)) but not in the absence of SCFA. However, in the in vitro experiments, the addition of 60 microM ethoxyzolamide had no significant effect on Jnet Ca2+. A reduction in pH of the intraruminal buffer in vivo from 6.8 to 5.4 led to significant increases in the net absorption rates of Ca2+ and Mg2+ ions, an effect which was duplicated for Ca2+ in preliminary in vitro experiments in which the pH of the mucosal buffer was reduced from 7.4 to 5.4. This stimulatory effect was confined to Jms Ca2+ and Jnet Ca2+. Ussing chambers were also used to demonstrate that Jnet Ca2+ was reduced by a high transmural potential difference (PD), caused by voltage clamping, independently of the mucosal K+ concentration. Both unidirectional Ca2+ fluxes consisted of a PD-dependent and a K+-insensitive PD-independent component. The latter may be represented by a Ca2+/ 2H+ antiporter. It is postulated that SCFA, and to a lesser extent H2CO3, can stimulate Jms Ca2+ by activation of an apical Ca2+/2H+ antiporter through the provision of protons within the ruminal epithelial cell. A mild reduction in ruminal pH may also lead to a similar stimulation of this putative electroneutral exchange.

[Effect of strophanthin on the absorption of sodium and short chain fatty acids from the sheep rumen]. / Vliianie strofantina na vsasyvanie natriia i korotkotsepochechnykh zhirnykh kislot iz predzheludkov ovets.

In vivo, intraruminal strophantin administration suppressed the sodium absorption by up to 50% in the sheep reticulo-rumen. Absorption of the shortchain fatty acids was not affected. The findings suggest absence of a combined transport of sodium and short-chain fatty acids' anions in basic and lateral membranes of the reticulo-rumen apithelium in sheep.

[Effect of acetazolamide on sodium and potassium absorption in the sheep forestomach]. / Vliianie atsetazolamida na vsasyvanie natriia i kaliia iz predzheludkov ovets.

I.v. Acetazolamide administration suppressed the sodium and potassium absorption up to 90 and 100%, resp., in the sheep reticulo-rumen. Absorption of the short chain fatty acids was not affected. The findings suggest a sodium/potassium exchange processes occurring in the reticulo-rumen epithelium in ruminants.

Use of a breath test to determine the fate of swallowed fluids in cattle.

OBJECTIVE: To investigate a [13C]octanoic acid breath test as a means of detecting reticular groove contraction in cattle. ANIMALS: 19 adult dairy cows with fistulated rumen, 10 yearling bulls, and 6 yearling steers. PROCEDURE: Cows were given 200 mg of [13C]octanoic acid in the caudal portion of the rumen, reticulum, or omasum/abomasum through the reticulo-omasal orifice, or were given the same dose of label with a drench of water or sodium bicarbonate. Collected breath was analyzed for 13C in CO2 for up to 3 hours. Breath of yearlings was analyzed for 13C in CO2 over 20 minutes after drenching with 200 mg of [13C]octanoic acid with water or sodium chloride and after sucking 200 mg of [13C]octanoic acid with molasses and water. RESULTS: In cows, enrichment of 13C in breath CO2 peaked at 20 to 30 minutes after placement of [13C]octanoic acid through the orifice, compared with a lower peak at 60 and 90 minutes after placement in the reticulum and rumen, respectively. The maximal increase in enrichment after placement of [13C]octanoic acid in the reticulum did not overlap with the minimal increase when placed through the reticulo-omasal orifice. Enrichment values in cows after drenching were consistent with values obtained after direct placement of [13C]octanoic acid. In yearlings, the inclusion of sodium chloride in the drench greatly increased enrichment, compared with water, but enrichment was greatest after sucking of the molasses, water, and [13C]octanoic acid combination. CONCLUSIONS AND CLINICAL RELEVANCE: This breath test provides a simple, repeatable, nonradioactive, and noninvasive means of detecting the fate of swallowed fluids in cattle, thus revealing the route taken of orally administered therapeutic agents or nutrients.

[Determination of the passage of two markers in the gastrointestinal tract of steers using different evaluation methods: effect of the level of nutrition]. / Ermittlung der passage zweier marker im verdauungstrakt von ochsen mit hilfe unterschiedlicher schätzverfahren: Einfluss des ernährungsniveaus.

This study was conducted to investigate the effects of amount of intake of a mixed diet by steers on the passage of digesta through the reticulorumen and the whole digestive tract. Six ruminally cannulated steers received a mixed diet consisting on average of 43% grass silage, 25% maize silage, 30% concentrate and 2% mineral-vitamin mix in DM. The experimental design was a repeated 3 x 3 Latin square with 21 days periods. The diet was offered twice daily (07.00 and 19.00 h) at 1, 1.5 and 2 times of estimated maintenance energy requirements. At the beginning of each period, animals received pulse doses of Titanium(IV)-oxide (TiO2) per os and Cr-EDTA intraruminally. Following marker administration, faecal marker concentrations were determined over 120 h. Passage parameters were estimated by a mono-exponential and a bi-exponential model, by models including gamma age-dependency and by a model calculating total tract mean retention time as mean value of all points on the marker-excretion curve. Passage rate of TiO2 from the reticulorumen increased with higher intakes, whereas mean retention time in the whole tract decreased. In general, results of different models were consistent across intakes. Values for passage of TiO2 were in good agreement with those reported for small particles, when similar diets were fed to cattle. Rate of passage of Cr-EDTA from the reticulorumen increased with higher intakes, and mean retention times of Cr-EDTA in the reticulorumen and in the whole tract decreased. Differences between models with or without age dependency were greater for Cr-EDTA than for TiO2. Fit to Cr-EDTA excretion curves was not satisfactory for models with gamma age dependency. Irrespective of model and marker, passage from the reticulorumen accelerated markedly, whereas retention times in the reticulorumen and in the whole tract decreased, as intake increased from maintenance energy requirements to 1.5 times maintenance.