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Disturbances of extracellular potassium (K) homeostasis in calves with severe neonatal diarrhea have been studied extensively. Although total body depletion of this predominantly intracellular electrolyte is generally thought to occur in diarrheic calves, the mechanisms through which K depletion occurs are poorly understood. The aim of this study was to investigate how intracellular K homeostasis is affected by dehydration and acidemia, the 2 most important metabolic disturbances in calves with naturally occurring diarrhea. Twenty-seven calves with naturally occurring neonatal diarrhea, pronounced dehydration, and acidemia, and 2 groups of 10 healthy control calves were included in this study. Blood samples and muscle biopsies were obtained immediately before initiation of treatment (T0) and after complete rehydration and correction of acidemia (T1) from diarrheic calves. Blood samples were used to perform blood gas, blood biochemical, and hematological analyses and to determine K content in erythrocytes. Muscle biopsies were used to determine muscle tissue K content and tissue dry matter. Controls were used to determine values for erythrocyte and muscle tissue K content in healthy neonatal calves for comparison with diarrheic calves. As defined by the inclusion criteria, diarrheic calves showed pronounced acidemia and dehydration at T0. Mean muscle tissue K content and tissue dry matter remained unchanged between sampling times and did not differ from values measured in healthy control calves. Erythrocyte K content increased from 73.63 ± 13.73 to 77.64 ± 15.97 mmol/L (±standard deviation) but was associated with a concomitant decline in erythrocyte volume. Values measured at both sampling times in diarrheic calves did not differ from erythrocyte K measured in healthy control calves. The plasma K concentration (median [interquartile range]) decreased from 5.44 [4.76-6.17] to 4.16 [3.99-4.31] mmol/L between T0 and T1. Although changes in plasma [K] were associated with the degree of dehydration, neither dehydration nor acidemia was associated with changes of K content in muscle tissue or erythrocytes. In conclusion, severe dehydration and acidemia in diarrheic calves were not associated with notable changes in K content of muscle tissue or erythrocytes. These results do not support the concept of pronounced K depletion occurring in calves with neonatal diarrhea. Erythrocytes are a poor surrogate tissue in which to measure changes of intracellular K content in diarrheic calves because of concomitant changes in erythrocyte volume that complicate the interpretation of results.

Magnesium (Mg2+) is an essential mineral without known specific regulatory mechanisms. In ruminants, plasma Mg2+concentration depends primarily on the balance between Mg2+absorption and Mg2+excretion. The primary site of Mg2+absorption is the rumen, where Mg2+is apically absorbed by both potential-dependent and potential-independent uptake mechanisms, reflecting involvement of ion channels and electroneutral transporters, respectively. Transport is energised in a secondary active manner by a basolateral Na+/Mg2+exchanger. Ruminal transport of Mg2+is significantly influenced by a variety of factors such as high K+concentration, sudden increases of ammonia, pH, and the concentration of SCFA. Impaired Mg2+absorption in the rumen is not compensated for by increased transport in the small or large intestine. While renal excretion can be adjusted to compensate precisely for any surplus in Mg2+uptake, a shortage in dietary Mg2+cannot be compensated for either via skeletal mobilisation of Mg2+or via up-regulation of ruminal absorption. In such situations, hypomagnesaemia will lead to decrease of a Mg2+in the cerebrospinal fluid and clinical manifestations of tetany. Improved knowledge concerning the factors governing Mg2+homeostasis will allow reliable recommendations for an adequate Mg2+intake and for the avoidance of possible disturbances. Future research should clarify the molecular identity of the suggested Mg2+transport proteins and the regulatory mechanisms controlling renal Mg excretion as parameters influencing Mg2+homeostasis.

High-performing dairy cows require a particular composition of nutritional ingredients, adapted to their individual requirements and depending on their production status. The optimal dimensioning of minerals in the diet, one being potassium, is indispensable for the prevention of imbalances. Potassium balance in cows is the result of potassium intake, distribution in the organism, and excretion, and it is closely related to glucose and electrolyte metabolism. In this paper, we present a dynamical model for potassium balance in lactating and nonlactating dairy cows based on ordinary differential equations. Parameter values were obtained from clinical trial data and from the literature. To verify the consistency of the model, we present simulation outcomes for 3 different scenarios: potassium balance in (1) nonlactating cows with varying feed intake, (2) nonlactating cows with varying potassium fraction in the diet, and (3) lactating cows with varying milk production levels. The results give insights into the short- and long-term potassium metabolism, providing an important step toward the understanding of the potassium network, the design of prophylactic feed additives, and possible treatment strategies.

Hypersecretion of chloride can cause diarrhea, a disease frequently occurring in young pigs, particularly around weaning. We investigated the contribution of different channels to intestinal Cl− secretion as influenced by age and weaning. Jejunal and colonic epithelia from 4-month-old pigs and 4-week-old piglets were incubated in Ussing chambers and stimulated by carbachol and forskolin. Changes in short-circuit currents were taken as measure of electrogenic net Cl− secretion. DIDS or NPPB served to inhibit Ca-activated Cl−-channels and outwardly rectifying Cl−-channels (ORCC) or cystic fibrosis transmembrane regulator (CFTR), respectively. Depolarizing the basolateral membrane allowed to examine the influence of K+-channels on Cl− secretion. Forskolin-stimulated Cl− secretion was mediated by CFTR. ORCC were not involved. Carbachol-induced Cl− secretion could be ascribed to an enhanced driving force due to the opening of K+-channels, whereas Ca-dependent Cl− channels seemed not to be involved. In jejunum, piglets showed higher Cl− secretion than pigs. Two days after weaning forskolin induced an I sc overshoot and a faster increase in G t. In colon, Cl− secretion was neither influenced by age nor by weaning. The data suggest a disposition of porcine jejunum for a higher Cl− secretion in young and freshly weaned piglets, which might be a natural defense mechanism as well as a predisposing factor for diarrhea.

Most cystic fibrosis (CF) patients show an exocrine pancreatic insufficiency that results in lower enzyme and bicarbonate secretion. To test whether an altered function of nutrient transporters might additionally attribute to the lower bodyweight of CF patients we investigated electrogenic absorption of alanine, glycyl-glutamine, glucose and the effect of pH on nutrient absorption by Ussing chambers in a CF mouse model carrying the Cftr TgH(neoim)Hgu mutation. The electrogenic transport of all three nutrients was similar between the D2.129P2(CF/3)-Cftr TgH(neoim)Hgu congenic strain and DBA/2J mice as well as between the B6.129P2(CF/3)-Cftr TgH(neoim)Hgu congenic strain and C57BL/6J mice. This indicates that the Cftr TgH(neoim)Hgu mutation does not affect the electrogenic absorption of alanine, glycyl-glutamine and glucose. In contrast, electrogenic nutrient absorption was reduced in the CF/1-Cftr TgH(neoim)Hgu and CF/3-Cftr TgH(neoim)Hgu inbred strains compared to the HsdOla:MF1, D2.129P2(CF/3)-Cftr TgH(neoim)Hgu and B6.129P2(CF/3)-Cftr TgH(neoim)Hgu strains, whereas no difference was found among the wild-type strains. This indicates that not the Cftr TgH(neoim)Hgu mutation but differences in the genetic background of the CF/1-Cftr TgH(neoim)Hgu and CF/3-Cftr TgH(neoim)Hgu strains compared to HsdOla:MF1, D2.129P2(CF/3)-Cftr TgH(neoim)Hgu and B6.129P2(CF/3)-Cftr TgH(neoim)Hgu strains are associated with the differences in electrogenic nutrient absorption. The electrogenic absorption of alanine, glycyl-glutamine and glucose was not influenced by an acidic pH (5.4) compared to absorption at pH 7.4.

In ruminants, more than 50% of overall gastrointestinal Ca absorption can occur preintestinally, and the anions of orally applied Ca salts are thought to play an important role in stimulating ruminal Ca absorption. This assumption is based mainly on ion-exchange studies that have used gluconate as the control anion, which may bind Ca2+ ions and interfere with treatment effects. In the present study, we investigated the distinct effects of different anions on Ca absorption across the sheep rumen and on the concentration of free Ca2+ ions ([Ca2+]ion). We showed that gluconate, sulfate, and short-chain fatty acids (SCFA) remarkably reduced [Ca2+]ion in buffer solutions. Nevertheless, increasing the Cl or SCFA concentration by 60 mM stimulated net ruminal Ca absorption 5- to 7-fold, but these effects could be antagonized by gluconate. Therefore, ion-exchange experiments must be (re)evaluated very carefully, because changes in [Ca2+]ion in the presence of gluconate, sulfate, or SCFA not only might entail an underestimation of Ca flux rates, but also might have effects on other cellular pathways that are Ca2+ dependent. Concerning the optimal Ca supply for dairy cows, the present study suggests that CaCl2 formulations and Ca salts of the SCFA stimulate Ca absorption across the rumen wall and are beneficial in preventing or correcting a Ca deficiency.

The quantitative role of different segments of the gastrointestinal tract for Ca absorption, the respective mechanisms, and their regulation are not fully identified for ruminants, that is, cattle. In different in vitro experiments the forestomach wall has been demonstrated to be a major site for active Ca absorption in sheep and goats. In order to further clarify the role of the bovine rumen for Ca transport with special attention to luminal Ca concentrations, its ionic form, and pH, electrophysiological and unidirectional flux rate measurements were performed with isolated bovine rumen epithelial tissues. For Ca flux studies (Jms, Jsm) in vitro Ussing chamber technique was applied. Standard RT-PCR method was used to characterize TRPV6 and PMCA1 as potential contributors to transepithelial active Ca transport. At Ca concentrations of 1.2 mmol L � 1 on both sides of the tissues, Jms were higher than Jsm resulting under some conditions in significant Ca net flux rates (Jnet), indicating the presence of active Ca transport. In the absence of an electrical gradient, Jnet could significantly be stimulated in the presence of luminal short-chain fatty acids (SCFAs). Increasing the luminal Ca concentrations up to 11.2 mmol L � 1 resulted in significant increases in Jms without influencing Jsm. Providing Ca in its form as respective chloride, formate, or propionate salts there was no significant effect on Jms. No transcripts specific for Ca channel TRPV6 could be demonstrated. Our results indicate different mechanisms for Ca absorption in bovine rumen as compared with those usually described for the small intestines.

High potassium diets lead to an inverse regulation of sodium and magnesium absorption in ruminants, suggesting some form of cross talk. Previous Ussing chamber experiments have demonstrated a divalent sensitive Na+ conductance in the apical membrane of ruminal epithelium. Using patch-clamped ruminal epithelial cells, we could observe a divalent sensitive, nonselective cation conductance (NSCC) with K+ permeability > Cs+ permeability > Na+ permeability. Conductance increased and rectification decreased when either Mg2+ or both Ca2+ and Mg2+ were removed from the internal or external solution or both. The conductance could be blocked by Ba2+, but not by tetraethylammonium (TEA). Subsequently, we studied this conductance measured as short-circuit current ( Isc) in Ussing chambers. Forskolin, IBMX, and theophylline are known to block both Isc and Na transport across ruminal epithelium in the presence of divalent cations. When the NSCC was stimulated by removing mucosal calcium, an initial decrease in Isc was followed by a subsequent increase. The cAMP-mediated increase in Isc was reduced by low serosal Na+ and serosal addition of imipramine or serosal amiloride and depended on the availability of mucosal magnesium. Luminal amiloride had no effect. Flux studies showed that low serosal Na+ reduced 28Mg fluxes from mucosal to serosal. The data suggest that cAMP stimulates basolateral Na+/Mg2+ exchange, reducing cytosolic Mg. This increases sodium uptake through a magnesium-sensitive NSCC in the apical membrane. Likewise, the reduction in magnesium uptake that follows ingestion of high potassium fodder may facilitate sodium absorption, as observed in studies of ruminal osmoregulation. Possibly, grass tetany (hypomagnesemia) is a side effect of this useful mechanism.

The left displacement of the abomasum (LDA) is a common disease in periparturient dairy cows. Plasma β-hydroxybutyrate (BHBA) levels above the reference range are regarded as risk factors for the occurrence of LDA. Additionally, hypokalemia and hypocalcemia have been observed in LDA cows. The aim of the present study was to characterize the membrane potential and the slow waves in abomasal smooth muscle cells by a microelectrode technique and to determine possible effects of BHBA and of various calcium and potassium concentrations on the membrane potential. Subsequently, the results obtained by the microelectrode technique were combined with in vitro motility experiments of abomasal smooth muscles. Strips of the abomasal circular smooth muscles were prepared and incubated in different buffer solutions. For the microelectrode technique, healthy bulls and cows that underwent surgery for LDA were sampled. These measurements showed a frequency of the slow waves between 3.5 and 10.9 per minute (for amplitudes ≥ 3mV) and between 0.6 and 4.5 per minute (for amplitudes ≥ 5mV). The frequency of contractions (1.8 to 3.1 per minute) were in the same order as the frequency of the slow waves with amplitudes ≥ 5 mV. Blocking potassium conductance with barium chloride induced a depolarization of the basal membrane potential (from -43±2.9 to -37±4.1mV; mean ± standard error of the mean) without affecting the frequency or the height of the slow waves. The reduction in the potassium concentration from 5.4 to 2 mmol/L resulted in a nominal decrease in the activity of contractions (from 22.2 to 18.6 mN/min). The subsequent addition of 1 mmol of KCl/L induced a nominal increase in contraction activity (from 18.6 to 25.7 mN/min). An effect of BHBA (5 mmol/L) could not be demonstrated, neither on the electric nor on the motility parameters. A simulated hypocalcemia (1.2 mmol/L total, 0.9 mmol/L ionized Ca) did not change slow waves and motility. In conclusion, changes in membrane potential ≥ 5 mV correlated with contractions of abomasal muscles. Hypokalemic conditions may reduce abomasal contraction activity via an effect on the membrane potential. An exclusive increase in BHBA or a slight hypocalcemia (0.9 mmol/L ionized Ca) had no effects on the motility of healthy abomasal smooth muscle cells. A prolonged or severe exposition of muscles to a combination of low Ca and high BHBA concentrations might nevertheless be able to affect abomasal motility.

Secretion of saliva as well as absorptive and secretory processes across forestomach epithelia ensures an optimal environment for microbial digestion in the forestomachs. Daily salivary secretion of sodium (Na+) exceeds the amount found in plasma by a factor of 2 to 3, while the secretion of bicarbonate (HCO3-) is 6 to 8 times higher than the amount of HCO3- in the total extracellular space. This implies a need for efficient absorptive mechanisms across forestomach epithelia to allow for an early recycling. While Na+ is absorbed from all forestomachs via Na+/H+ exchange and a non-selective cation channel that shows increased conductance at low concentrations of Mg2+, Ca2+ or H+ in the luminal microclima and at low intracellular Mg2+, HCO3- is secreted by the rumen for the buffering of ingesta but absorbed by the omasum to prevent liberation of CO2 in the abomasum. Fermentation provides short chain fatty acids and ammonia (NH3) that have to be absorbed both to meet nutrient requirements and maintain ruminal homeostasis of pH and osmolarity. The rumen is an important location for the absorption of essential minerals such as Mg2+ from the diet. Other ions can be absorbed, if delivered in sufficient amounts (Ca2+, Pi, K+, Cl- and NH4+). Although the presence of transport mechanisms for these electrolytes has been described earlier, our knowledge about their nature, regulation and crosstalk has increased greatly in the last years. New transport pathways have recently been added to our picture of epithelial transport across rumen and omasum, including an apical non-selective cation conductance, a basolateral anion conductance, an apical H+-ATPase, differently expressed anion exchangers and monocarboxylate transporters.

Background A major boost to cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original Cftr TgH(neoim)Hgu mouse model with a divergent genetic background (129/Sv, C57BL/6, HsdOla:MF1) two inbred mutant mouse strains CF/1-Cftr TgH(neoim)Hgu and CF/3-Cftr TgH(neoim)Hgu had been generated using strict brother × sister mating. CF/1-Cftr TgH(neoim)Hgu and CF/3-Cftr TgH(neoim)Hgu mice were fertile and showed normal growth and lifespan. In this work the Cftr TgH(neoim)Hgu insertional mutation was backcrossed from CF/3-Cftr TgH(neoim)Hgu onto the inbred backgrounds C57BL/6J and DBA/2J generating congenic animals in order to clarify the differential impact of the Cftr mutation and the genetic background on the disease phenotype of the cystic fibrosis mutant mice. Clinical and electrophysiological features of the two congenic strains were compared with those of CF/1-Cftr TgH(neoim)Hgu and CF/3-Cftr TgH(neoim)Hgu and wild type controls. Results Under the standardized housing conditions of the animal facility, the four mouse strains CF/1-Cftr TgH(neoim)Hgu , CF/3-Cftr TgH(neoim)Hgu , D2.129P2(CF/3)-Cftr TgH(neoim)Hgu and B6.129P2(CF/3)-Cftr TgH(neoim)Hgu exhibited normal life expectancy. Growth of congenic cystic fibrosis mice was comparable with that of wild type controls. All mice but D2.129P2(CF/3)-Cftr TgH(neoim)Hgu females were fertile. Short circuit current measurements revealed characteristic response profiles of the HsdOla:MF1, DBA/2J and C57BL/6J backgrounds in nose, ileum and colon. All cystic fibrosis mouse lines showed the disease-typical hyperresponsiveness to amiloride in the respiratory epithelium. The mean chloride secretory responses to carbachol or forskolin were 15–100% of those of the cognate wild type control animals. Conclusion The amelioration of the clinical features and of the basic defect that had emerged during the generation of CF/3-Cftr TgH(neoim)Hgu mice was retained in the congenic mice indicating that the Cftr linkage group or other loci shared between the inbred strains contain(s) the major modifier(s) of attenuation of cystic fibrosis symptoms.

Oesophagostomum dentatum, one of the most common nematodes in pigs, causes the formation of subepithelial granuloma in the large intestine. To investigate possible changes in epithelial function or response during the infection we incubated epithelia of pig proximal colon in Ussing chambers at different days post infectionem (p.i.). Transepithelial conductance, gt, and the Cl flux from serosal to mucosal, JClsm, were increased on day 2 p.i., when the nematodes penetrate the epithelium of the large intestine, and declined toward control levels thereafter. Histamine, PGE2 and carbachol caused transient increases in short circuit current, Isc, and conductance that could partly be attributed to a higher JClsm. The Isc responses were highest on the days of nematode penetration in or out of the epithelium (days 2 and 14 p.i.) and did decline on day 7 p.i. during the histotropic development of the parasite. This reduced epithelial reaction on day 7 p.i. might be an adaptation to secretory stimuli released from the inflammatory cells in the intestinal wall or might reflect modulation by the parasite and could be responsible for the absence of marked clinical signs during the infection.

Short chain fatty acids (SCFAs) and CO2 have been shown to stimulate net Mg2+ efflux from the isolated reticulorumen in vivo. To investigate the underlying mechanisms of Mg2+ transport we performed Ussing chamber and microelectrode experiments and measured 28Mg2+ fluxes across sheep rumen epithelium in vitro. In the presence of SCFAs mucosal-to-serosal Mg2+ flux (Jm-sMg) amounted to 82.3 +/- 7.8 nmol cm-2 h-1 and serosal-to-mucosal Mg2+ flux (Js-mMg) to 3.2 +/- 0.7 nmol cm-2 h-1. Replacing SCFAs with gluconate caused a 50% reduction of Jm-sMg, whereas Js-mMg was not affected. Among the SCFAs, n-butyrate was more effective in stimulating Jm-sMg than acetate, propionate or iso-butyrate. Eliminating HCO3(-)-CO2 from SCFA-containing solutions did not affect Mg2+ fluxes, whereas the same replacement in SCFA-free solutions led to a further reduction in Jm-sMg. Jm-sMg decreased after the addition of ethoxyzolamide to SCFA-free, bicarbonate buffered solutions. Decreasing mucosal pH from 6.4 to 5.4 increased Jm-sMg in SCFA-free, bicarbonate buffered solutions. SCFAs had no effect on the apical membrane potential of rumen epithelial cells. The experiments show that both SCFAs and CO2 stimulate Mg2+ transport through an increase in Jm-sMg, most probably via stimulation of a Mg(2+)-2H+ exchange mechanism. SCFAs may have additional metabolic effects on Mg2+ transport.