Der erfolgreiche Lehrbuchklassiker für Studierende der Agrarwissenschaften und Veterinärmedizin, aktualisiert und erweitert! In der 16. Auflage wird verstärkt die Entwicklung von Krankheiten im Zusammenhang mit den anatomischen und physiologischen Gegebenheiten behandelt. Neben der Anatomie und Physiologie der Nutztiere einschließlich des Geflügels wird jetzt auch die des Hundes vertiefend dargelegt. Komplexe Sachverhalte werden durch Illustrationen leichter verständlich. Aus dem Inhalt: Zelle und Gewebe Skelettsystem, Zähne und Skelettmuskelsystem Blut und Infektionsabwehr Herz, Kreislaufsystem und Gefäße Atmungsapparat und Verdauungsapparat Energiehaushalt und Wärmehaushalt Harnorgane, Geschlechtsorgane und Milchdrüse Haut und Anhangsgebilde Nervensystem, Sinnesorgane, Endokrines System Säure-Basen-Haushalt (neu in der 16. Auflage) Im Anhang: Bezeichnungen, SI-Einheiten und Umrechnungen, Abkürzungen und Begriffserklärungen lateinischer und griechischer Termini.

Chronic inflammatory enteropathy (CIE) in dogs, a spontaneous model of human inflammatory bowel disease (IBD), is associated with a high rate of cobalamin deficiency. The etiology of hypocobalaminemia in human IBD and canine CIE remains unknown, and compromised intestinal uptake of cobalamin resulting from ileal cobalamin receptor deficiency has been proposed as a possible cause. Here, we evaluated the intestinal expression of the cobalamin receptor subunits, amnionless (AMN) and cubilin (CUBN), and the basolateral efflux transporter multi-drug resistance protein 1 (MRP1) in 22 dogs with CIE in comparison to healthy dogs. Epithelial CUBN and AMN levels were quantified by confocal laser scanning microscopy using immunohistochemistry in endoscopic ileal biopsies from dogs with (i) CIE and normocobalaminemia, (ii) CIE and suboptimal serum cobalamin status, (iii) CIE and severe hypocobalaminemia, and (iv) healthy controls. CUBN and MRP1 expression was quantified by RT-qPCR. Receptor expression was evaluated for correlation with clinical patient data. Ileal mucosal protein levels of AMN and CUBN as well as mRNA levels of CUBN and MRP1 were significantly increased in dogs with CIE compared to healthy controls. Ileal cobalamin receptor expression was positively correlated with age, clinical disease activity index (CCECAI) score, and lacteal dilation in the ileum, inversely correlated with serum folate concentrations, but was not associated with serum cobalamin concentrations. Cobalamin receptor downregulation does not appear to be the primary cause of hypocobalaminemia in canine CIE. In dogs of older age with severe clinical signs and/or microscopic intestinal lesions, intestinal cobalamin receptor upregulation is proposed as a mechanism to compensate for CIE-associated hypocobalaminemia. These results support oral supplementation strategies in hypocobalaminemic CIE patients.

An optimal fermentation process in the forestomach is pivotal for the wellbeing and performance of ruminants. Complex carbohydrates are broken down into short-chain fatty acids (SCFA) which form the major energy source for the animal. A strong interrelationship of this process with intraruminal pH and redox potential (Eh) exists. These parameters can be measured with intraruminal sensors, but the interpretation of the measurements, especially of Eh, and their meaning for intraruminal homeostasis is not completely clear. In this study, factors influencing intraruminal Eh were elucidated. We hypothesised that intraruminal Eh is influenced by the fermentation process as such, but not by its end products SCFA. We measured Eh and pH in ruminal fluid from fasting cannulated sheep after the addition of 0.06 m Na-acetate, -propionate, -butyrate or glucose in vitro. Furthermore, we assessed the interrelation of pH and Eh. Basal Eh and pH values were -120 ± 41 mV and 7.0 ± 0.3, respectively, in native ruminal fluid in vitro. While the addition of SCFA did not induce any changes, glucose addition caused a significant decrease in both pH and Eh compared to the values before the addition (paired Student's t-test, p < 0.05). We attribute the decrease in Eh to an increased production of H2 in the process of generating SCFA, predominantly acetate. By titrating both native and particle-free ruminal fluid to more acidic and basic pH values (4.5-8.5), we found a non-linear inverse correlation of pH and Eh, counteracting the H2 -driven decrease of Eh during fermentation. Thus, the intraruminal Eh is influenced by pH and H2 output during SCFA formation. The opposed character of these factors stabilises the intraruminal homeostasis which might help maintain symbiotic microbiota in the rumen. Understanding, monitoring, and supporting this system will be an essential part of modern cattle production.

The large intestinal epithelium is confronted with the necessity to adapt quickly to varying levels of oxygenation. In contrast to other tissues, it meets this requirement successfully and remains unharmed during (limited) hypoxic periods. The large intestine is also the site of bacterial fermentation producing short-chain fatty acids (SCFA). Amongst these SCFA, butyrate has been reported to ameliorate many pathological conditions. Thus, we hypothesized that butyrate protects the colonocytes from hypoxic damage. We used isolated porcine colon epithelium mounted in Ussing chambers, incubated it with or without butyrate and simulated hypoxia by changing the gassing regime to test this hypothesis. We found an increase in transepithelial conductance and a decrease in short-circuit current across the epithelia when simulating hypoxia for more than 30 min. Incubation with 50 mM butyrate significantly ameliorated these changes to the epithelial integrity. In order to characterize the protective mechanism, we compared the effects of butyrate to those of iso-butyrate and propionate. These two SCFAs exerted similar effects to butyrate. Therefore, we propose that the protective effect of butyrate on colon epithelium under hypoxia is not (only) based on its nutritive function, but rather on the intracellular signaling effects of SCFA.

We hypothesized that, due to the high pH of this compartment, the reticulum epithelium displays particular features in the transport of short-chain fatty acids (SCFA). Ovine reticulum epithelium was incubated in Ussing chambers using a bicarbonate-free buffer solution containing butyrate (20 mmol L−1). p-hydroxymercuribenzoic acid (pHMB), 5-(N-Ethyl-N-isopropyl)amiloride (EIPA), or ouabain were added to the buffer solution as inhibitors of monocarboxylate transporters, sodium-proton-exchangers, or the Na+/K+-ATPase, respectively. The short-circuit current (Isc) and transepithelial conductance (Gt) were monitored continuously while the flux rates of 14C-labelled butyrate were measured in the mucosal-to-serosal (Jmsbut) or serosal-to-mucosal direction (Jsmbut). Under control conditions, the mean values of Isc and Gt amounted to 2.54 ± 0.46 µEq cm−2 h−1 and 6.02 ± 3.3 mS cm−2, respectively. Jmsbut was 2.1 ± 1.01 µmol cm−2 h−1 on average and about twice as high as Jsmbut. Incubation with ouabain reduced Jmsbut, while Jsmbut was not affected. The serosal addition of EIPA did not affect Jmsbut but reduced Jsmbut by about 10%. The addition of pHMB to the mucosal or serosal solution reduced Jmsbut but had no effect on Jsmbut. Mucosally applied pHMB provoked a transient increase in the Isc. The serosal pHMB sharply reduced Isc. Our results demonstrate that butyrate can be effectively transported across the reticulum epithelium. The mechanisms involved in this absorption differ from those known from the rumen epithelium.

High amounts of short-chain fatty acids (SCFAs) occur in the ovine rumen and constitute the animal's main energy source. However, they lead to an acidification of the ruminal epithelium. Therefore, effective intracellular pH (pHi ) regulation by transport proteins like monocarboxylate transporter 1 (MCT1) and Na+ /H+ exchangers (NHEs) is pivotal to ruminants to avoid epithelial damage. SCFAs might function not only as nutrients but also as signalling molecules by activating free fatty acid receptors (FFARs) in the ruminal epithelium and thus influence pHi regulation. FFARs work as nutrient sensors, transducing their information by modulating cyclic adenosine monophosphate (cAMP) levels. We hypothesized that (FFAR-modulated) decreases in cAMP levels stimulate the activity of MCT1 and NHEs in the ruminal epithelium of sheep. We detected two FFARs (GPR109A and FFAR2) immunohistochemically in the ovine ruminal epithelium. Administration of 10 mM butyrate to Ussing chamber-mounted epithelia provoked a significant reduction in intraepithelial cAMP levels. However, application of the GPR109A agonist niacin did not affect cAMP levels. MCT1 activity was analysed by measuring transepithelial 14 C-acetate fluxes, which were not inhibited by forskolin-induced increased cAMP levels. The recovery of pHi after acidification was assessed as an indicator of NHE activity in primary cultured ruminal epithelial cells. Recovery was significantly reduced when cells with increased cAMP levels were subjected to the NHE inhibitor 5-(N-ethyl-N-isopropyl)-amiloride (10 µM). Nonetheless, with augmented cAMP levels alone, NHE activity tended to decline. We hypothesize that modulation of cAMP levels by butyrate is accomplished by FFAR2 activation, regulating NHE activity for pHi homoeostasis at least in part.

The reticulorumen, as the main fermentation site of ruminants, delivers energy in the form of short-chain fatty acids (SCFA) for both the animal as well as the ruminal wall. By absorbing these SCFA, the ruminal epithelium plays a major role in the maintenance of intraruminal and intraepithelial acid-base homoeostasis as well as the balance of osmolarity. It takes up SCFAviaseveral pathways which additionally lead to either a reduction of protons in the ruminal lumen or the secretion of bicarbonate, ultimately buffering the ruminal content effectively. Nutrition of the epithelium itself is achieved by catabolism of the SCFA, especially butyrate. Catabolism of SCFA also helps to maintain a concentration gradient across the epithelium to ensure efficient SCFA uptake and stability of the epithelial osmolarity. Furthermore, the ruminal epithelium forms a tight barrier against pathogens, endotoxins or biogenic amines, which may emerge from ruminal microorganisms and feed. Under physiological conditions, it reduces toxin uptake to a minimum. Moreover, the epithelium seems to have the ability to degrade biogenic amines like histamine. Nonetheless, in high performance production animals like dairy cattle, the reticulorumen is confronted with large amounts of rapidly fermentable carbohydrates. This may push the epithelium to its limits, even though it possesses a great capacity to adapt to varying feeding conditions. If the epithelial limit is exceeded, increasing amounts of SCFA lead to an acidotic imbalance that provokes epithelial damage and thereby elevates the entrance of pathogens and other potentially harmful substances into the animal's body. Hence, the ruminal epithelium lays the foundation for the animal's health, and in order to ensure longevity and high performance of ruminant farm animals, it should never be overburdened.

The intestinal epithelium is able to adapt to varying blood flow and, thus, oxygen availability. Still, the adaptation fails under pathologic situations. A better understanding of the mechanisms underlying the epithelial adaptation to hypoxia could help to improve the therapeutic approach. We hypothesized that the short-term adaptation to hypoxia is mediated via AMP-activated protein kinase (AMPK) and that it is coupled to the long-term adaptation by a common regulation mechanism, the HIF-hydroxylase enzymes. Further, we hypothesized the transepithelial transport of glucose to be part of this short-term adaptation. We conducted Ussing chamber studies using isolated lagomorph jejunum epithelium and cell culture experiments with CaCo-2 cells. The epithelia and cells were incubated under 100% and 21% O2, respectively, with the panhydroxylase inhibitor dimethyloxalylglycine (DMOG) or under 1% O2. We showed an activation of AMPK under hypoxia and after incubation with DMOG by Western blot. This could be related to functional effects like an impairment of Na+-coupled glucose transport. Inhibitor studies revealed a recruitment of glucose transporter 1 under hypoxia, but not after incubation with DMOG. Summing up, we showed an influence of hydroxylase enzymes on AMPK activity and similarities between hypoxia and the effects of hydroxylase inhibition on functional changes.

Der erfolgreiche Lehrbuch-Klassiker für Studierende der Agrarwissenschaften und Veterinärmedizin jetzt im neuen Format! Für die 15. Auflage wurden die Illustrationen komplett neu gestaltet und der Text didaktisch überarbeitet, schwierige Sachverhalte wurden verständlich illustriert. Neu aufgenommen wurden die Anatomie und Physiologie des Geflügels. Im Anhang: Bezeichnungen, SI-Einheiten und Umrechnungen, Abkürzungen und Begriffserklärungen lateinischer und griechischer Termini sowie ein umfangreiches Stichwortverzeichnis! Kompakt, übersichtlich und leicht verständlich - ideal zum Lernen und für die Prüfungsvorbereitung.

In horses, strangulation of the intestine often leads to hypoxia in jejunal loops resulting in irreversible lesions of the epithelium. This contrasts with the common view that this epithelium is able to adapt to different oxygenation conditions well. To get a better understanding of the pathomechanisms of hypoxic injury, our study aimed to investigate functional adaptation processes in equine jejunum epithelium. Therefore, we incubated jejunal epithelia of horses in Ussing chambers and simulated hypoxia by gassing with 1% oxygen or by incubation with the chemical hypoxia-mimetic CoCl2. Because glucose is an important fuel for the cells' metabolism and thus its import is even more important under hypoxia, we assessed glucose transport mechanisms by measuring its electrogenic transport via sodium-coupled glucose transporter (SGLT1) and the transepithelial transport of 14C-glucose and 14C-ortho-methyl-glucose (OMG) as well as mRNA expression of glucose transporters. We could show that electrogenic uptake of glucose into the epithelium via SGLT1 was downregulated under hypoxia as was the mRNA expression of SGLT1 after CoCl2 incubation. However, transepithelial transport of glucose and OMG was not altered. Thus, there must be alterations in transepithelial transport mechanisms securing an energy-independent uptake of glucose into the epithelial cells. We believe that these changes are probably mediated on protein level as we found only effects of prolonged CoCl2 incubation on gene expression while the functional effects observed occurred much earlier.

Continuous cell lines have become indispensable tools that have enabled investigations into cellular mechanisms by increasing experimental reproducibility and sample availability, and decreasing the use of experimental animals. To facilitate studies of epithelial barrier function of the porcine colon, we aimed to establish an epithelial cell line with an extended replicative capacity. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen (SV40 TAg). We established a clonal epithelial cell line, referred to as PoCo83-3, that stably expressed the SV40 TAg, verified at mRNA and protein levels. PoCo83-3 showed epithelial cell-specific features, such as cobblestone-like morphology, dome structure formation, the presence of apical microvilli, and the expression of keratin 18, E-cadherin and the tight junction-associated proteins zonula occludens-1, occludin, and claudin-1. To validate PoCo83-3 as an in vitro model in epithelial barrier research, proinflammatory cytokine-inducible alterations in barrier integrity were demonstrated by incubating the cells with TNF-α and IFN-γ for 48 h. These cytokine treatments promoted a decreased transepithelial electrical resistance. In summary, PoCo83-3 exhibited an extended life span and a differentiated phenotype while maintaining epithelial characteristics. Based on these results, we present this cell line as a valuable in vitro model for investigations of epithelial barrier function in the porcine colon.

The present study pursued the question if an anion channel could be partly responsible for the transport of acetate in the isolated ovine ruminal epithelium. Using the Ussing chamber technique, changes of short-circuit current (Isc) induced by mucosal or serosal addition of acetate was investigated. To further evaluate the Isc changes induced by acetate the epithelia were preincubated with ouabain or 4,4′-diisothiocyano-2,2′-stilbenedisulfonic acid (DIDS). In addition, unidirectional flux rates of 14C-acetate were measured at different transepithelial potential difference (Vt). At Vt=0mV, acetate addition to the mucosal side of epithelia incubated in ouabain/DIDS free solution resulted in an Isc decrease, whereas application to the serosal side induced an Isc increase. Absolute Isc changes were significantly larger after serosal than after mucosal acetate addition. Ouabain pre-incubation abolished these side-specific differences. Pre-incubation with DIDS on the mucosal side inhibited the current induced by subsequent mucosal acetate addition in a voltage dependent manner, whereas serosal DIDS pre-incubation had no effect on acetate-induced changes of Isc. Mucosal-to-serosal acetate flux was partly voltage-dependent. The serosal-to-mucosal flux of acetate was not influenced by Vt. The asymmetric changes of Isc after acetate application and the Vt dependence of the DIDS inhibition on the mucosal side indicate a partly electrogenic transcellular permeation of acetate which includes an apically located acetate conductance. It is suggested that this conductance may be important for epithelial cell homeostasis.

The gastrointestinal epithelium possesses adaptation mechanisms to cope with huge variations in blood flow and subsequently oxygenation. Since sufficient energy supply is crucial under hypoxic conditions, glucose uptake especially must be regulated by these adaptation mechanisms. Therefore, we investigated glucose transport under hypoxic conditions. Jejunal epithelia of rabbits were incubated in Ussing chambers under short-circuit current conditions. Hypoxia was simulated by gassing with 1% O2 instead of 100% O2. The activity of sodium-coupled glucose transporter-1 (SGLT-1) was assessed by measuring the increase of short circuit current ( Isc) after the addition of 2 mM glucose to the mucosal buffer solution. We observed decreased activity of SGLT-1 after hypoxia compared with control conditions. To investigate underlying mechanisms, epithelia were exposed to agonists and antagonists of AMP-activated protein kinase (AMPK) before assessment of SGLT-1-mediated transport and the pAMPK/AMPK protein ratio. Preincubation with the antagonist restored SGLT-1 activity under hypoxic conditions to the level of control conditions, indicating an involvement of AMPK in the downregulation of SGLT-1 activity under hypoxia, which was confirmed in Western blot analysis of pAMPK/AMPK. Transepithelial flux studies using radioactively labeled glucose, ortho-methyl-glucose, fructose, and mannitol revealed no changes after hypoxic incubation. Therefore, we could exclude a decreased transepithelial glucose transport rate and increased paracellular conductance under hypoxia. In conclusion, our study hints at a decreased activity of SGLT-1 under hypoxic conditions in an AMPK-dependent manner. However, transepithelial transport of glucose is maintained. Therefore, we suggest other transport mechanisms, especially glucose transporter 1 and/or 2 to substitute SGLT-1 under hypoxia. NEW & NOTEWORTHY To our knowledge, this is the first approach to simulate hypoxia and study its effects in the jejunal epithelium using the Ussing chamber technique. We were able show that AMPK plays a role in the downregulation of SGLT-1 and that there seems to be an upregulation of other glucose transport mechanisms in the apical membrane of lagomorph jejunum epithelium under hypoxia, securing the epithelial energy supply and thus integrity.