Absorption of short-chain fatty acids,sodium and water from the forestomach of camels

In camelids the ventral parts of compartments 1 and 2 (C1/C2) and the total surface of compartment 3 of the forestomach are lined with tubular glands, whereas in ruminants the surface of the forestomach is composed entirely of stratified, squamous epithelium. Thus, differences in absorption rates between these foregut fermenters can be expected. In five camels C1/C2 was temporarily isolated, washed and filled with buffer solutions. Absorption of short-chain fatty acids (SCFA) and net absorption of sodium and water were estimated relative to Cr-ethylenediaminetetraacetic acid as a fluid marker. SCFA were extensively absorbed in the forestomach; clearance rates of SCFA with different chain lengths were equal. After lowering the pH of solutions SCFA absorption rates increased, but much less than the increase of the non-ionized fraction. Absorption of propionate was lower when acetate had been added. Findings suggest that most of the SCFA in camels are transported in the ionized form, most likely via an anion exchange mechanism. Net water absorption is closely related to net sodium absorption. Apparently water absorption results from an iso-osmotic process. Differences between absorption mechanisms of SCFA from the forestomach of camelids and ruminants are discussed.     

Histochemische,elektronenmikroskopische und biochemische Untersuchungen über die ATPasen im Vormagenepithel der Ziege

Zusammenfassung Am Epithel der drei Vormägen der Ziege wurden licht- und elektronenmikroskopische Untersuchungen über den Nachweis der Mg⁺⁺-, (Mg⁺⁺-Na⁺-K⁺)- und der Ca⁺⁺-aktivierbaren ATPasen durchgeführt. Bereits an lichtmikroskopischen Präparaten wurde festgestellt, daß sich deutliche Unterschiede in der Enzymaktivität der Mg⁺⁺- und Ca⁺⁺ stimulierbaren ATPasen sowohl für die einzelnen Epithelschichten als auch für die verschiedenen Bereiche der Vormägen nachweisen lassen; die tiefen Epithellagen reagieren stets stärker als die oberflächlichen. Ferner ist die in hohem Maße von der Inkubationszeit abhängige Reaktionsstärke im Epithel des Blättermagens größer als in der Lamina epithelialis des Netzmagens und vor allem in jener des Pansens. Bei dem elektronenmikroskopisch geführten Nachweis der ATPasen zeigt sich, daß die Reaktionsprodukte an der Außenseite der Zellmembranen liegen. Frei von Niederschlägen sind alle Zellmembranabschnitte, die sich an der Bildung der Desmosomen und Halbdesmosomen beteiligen sowie die basalen Abschnitte der Basalzellen.Im Hinblick auf die Verteilung und feinstrukturelle Lokalisation konnten keine Unterschiede zwischen den Mg⁺⁺-, Ca⁺⁺- und (Mg⁺⁺-Na⁺-K⁺)-aktivierbaren ATPasen beobachtet werden.In Anlehnung an die Methode nach Coleman u.a. (1967) wurden Zellmembranen von Pansenepithelzellen isoliert und an diesen biochemische Enzymbestimmungen durchgeführt. Die Ausbeute der gewonnenen Zellmembranen betrug, gemessen anhand der 5-Nukleotidase, dem Leitenzym für Plasmamembranen, gegenüber dem eingesetzten Gesamthomogenat 0,3%. Die mit diesem Verfahren durchgeführten biochemischen Enzymbestimmungen erbrachten den Nachweis, daß in den Plasmamembranen der Pansenepithelzellen neben der Mg⁺⁺- und Ca⁺⁺-aktivierbaren auch eine (Na⁺-K⁺)-abhängige Transport-ATPase vorkommt.

---

Histochemical, ultrastructural and biochemical studies of atpases of the goat forestomach epithelium

Summary Light- and electron microscopical studies were carried out to demonstrate ATPases activated by Mg⁺⁺, by (Mg⁺⁺-Na⁺-K⁺), and by Ca⁺⁺. Histological sections showed clear differences of the activity of Mg⁺⁺- and Ca⁺⁺-stimulated ATPases in the light microscope in different layers of the epithelium as well as in different areas of the forestomach. The deeper layers reacted more intensely than the superficial ones. The intensity of the reaction (which depends on incubation time) in the omasal epithelium was stronger than in the lamina epithelialis of the reticulum and much stronger than in the lamina epithelialis of the rumen. In the electron microscope, the reaction products of the ATPase appeared on the outer surface of cell membranes (plasmalemmata). No deposits of the reaction products were observed on those areas of the cell membranes, which are involved in the formation of desmosomes and semidesmosomes. The basal parts of the basal cells were also free from reaction products. As for the distribution and ultrastructural localisation of the deposits, no differences were observed among the ATPase stimulated by Mg⁺⁺, (Mg⁺⁺-Na⁺-K⁺), and Ca⁺⁺.Using the technique of Coleman et al. (1967), the cell membranes of ruminal epithelium were isolated. Biochemical assays of the enzymes were carried out. The quantity of cell membranes obtained was 0.3% of the whole homogenate, when compared with 5-nucleotidase, which is the typical enzyme of plasmalemmata. The biochemical enzyme assays showed that, besides Mg⁺⁺- and Ca⁺⁺-dependent ATPases, a (Na⁺-K⁺)-dependent transport ATPase exists in the cell membranes of ruminal epithelial cells.

Die cytochemischen Ergebnisse wurden auszugsweise auf dem Kongreß der Europäischen Vereinigung der Veterinäranatomen vom 9.–11. September 1968 in Belgrad vorgetragen.     

Cytochemische Untersuchungen über die alkalische Phosphatase im Vormagenepithel der Ziege

Zusammenfassung Mit licht- und elektronenmikroskopischen Methoden wurde die Verteilung und Aktivität der unspezifischen alkalischen Phosphatase im Epithel der drei Vormägen der Ziege untersucht und gleichzeitig der Einfluß verschiedener Fixiermittel und unterschiedlicher Vorfixierungs- und Inkubationszeiten auf das Ergebnis der Enzymreaktion geprüft. Es wurde festgestellt, daß die alkalische Phosphatase bei allen untersuchten Proben nur im Stratum corneum, Stratum granulosum und Stratum spinosum superf. vorkommt. Verschiedene Vorfixierungen der Proben entweder mit Formol-Kalzium (24 Std) oder Glutaraldehyd (2, 5, 30, 120 min) bzw. OsO₄ (2 oder 5 min) beeinflussen die Enzymverteilung und Reaktionsstärke nicht. Beim lichtmikroskopischen Nachweis wurde die maximale Reaktionsstärke bereits bei einer Inkubationszeit von 10 min bei Zimmertemperatur erreicht. Für den elektronenmikroskopischen Enzymnachweis war eine Inkubationszeit von 5 min bei 4° C am günstigsten. Die Reaktionsprodukte sind sowohl an den Zellmembranen als auch in Zellpartikeln lokalisiert. Die zellmembrangebundenen Reaktionsprodukte befinden sich bei allen mit Glutaraldehyd vorfixierten Proben an der äußeren Lamelle, bei kurzzeitiger Osmiumvorfixation (3 min) hingegen an der Innenseite der Plasmamembranen.

---

Cytochemical study of alkaline phosphatase activity in the goat forestomach epithelium

Summary Light and electron microscopic techniques were employed to study the distribution and intensity of the non-specific alkaline phosphatase activity in the epithelium of the three forestomachs of the goat. The effects on the enzyme reaction of different fixatives, prefixatives, and incubation times were determined.Alkaline phosphatase was found to be present only in the stratum corneum, stratum granulosum, and stratum spinosum superf. of each of the specimens. Different prefixation of the specimens, either by formol calcium (24 h), glutaraldehyde (2, 5, 30, 120 min), or osmium tetroxide (2 or 5 min) had no influence on the distribution of the enzyme and the intensity of its reaction. Maximal intensity of the reaction was obtained after an incubation period of 10 min at room temperature, as seen with the light microscope. To demonstrate the enzyme in sections in the electron microscope, an incubation period of 5 min at 4° C was found to be optimal. The products of the enzyme reaction were located on cell membranes and in cell particles. The membrane-bound reaction products in the specimens prefixed with glutaraldehyde were found on the outer surface of the plasma membrane; after a short prefixation with osmium tetroxide (3 min), they appeared on the inner surface of the plasma membrane.

Auszugsweise vorgetragen auf dem Kongreß der Europäischen Vereinigung der Veterinäranatomen vom 8.–10. September 1969 in Parma.     

Intake, ingesta retention, particle size distribution and digestibility in the hippopotamidae

Although several aspects of the digestive physiology of the hippopotamidae-non-ruminating foregut fermenters-have been described, ingesta kinetics and passage characteristics of these species are not well understood. The most outstanding feature of the hippo digestive physiology reported so far is the very long mean ingesta retention times (MRTs) measured by Foose [Foose, T., 1982. Trophic strategies of ruminant versus nonruminant ungulates. PhD dissertation, University of Chicago, Chicago.]. Since those data had been investigated with animals without water access, we intended to measure MRT in hippos which were allowed to enter water pools during the night. MRT parameters as well as dry matter (DM) digestibility were determined in four common (Hippopotamus amphibius) and four pygmy hippos (Hexaprotodon liberiensis) on two different diets each using cobalt ethylendiamintetraacetate (Co-EDTA) as a fluid, chromium (Cr)-mordanted fibre (<2 mm) as a particle and acid detergent lignin (ADL) as an internal digestibility marker. Four of the animals additionally received cerium (Ce)-mordanted fibres (2-10 mm) as particle markers. Total MRTs for fluids and particles ranged between 20-35 and 48-106 h in the common and between 13-39 and 32-107 h in the pygmy hippos. The difference between fluid and particle retention was greater than usually reported in ruminants. Excretion patterns of the markers differed from those usually observed in ruminants but resembled those reported for macropods (kangaroos), indicating a plug-flow reactor-like physiology in the hippo forestomach (FRST). This finding complements other described similarities between the macropod and the hippo forestomach. The measurements of larger particle retention profiles suggest that in the hippo, larger particles might be excreted either faster or at the same rate as smaller particles, indicating a general difference between ruminants and hippos with respect to differential particle retention. The digestive physiology of hippos is characterised by a generally low food intake, long ingesta retention times and dry matter digestibilities lower than reported in ruminants. Moderate digestibilities in spite of long retention times might be the result of the generally high average ingesta particle size in hippos. The comparatively easy management of pygmy hippos, together with the significant correlations between food intake, MRT and digestibility in the pygmy hippos of this study, recommends this species for further studies on the interplay of these parameters in herbivore digestive physiology.     

袖状胃切除术Wistar大鼠模型建立及初步观察

目的探讨袖带胃切除术对Wistar大鼠体质量、大鼠前胃黏膜及黏膜肌层厚度的影响,为进一步研究袖状胃切除术的减重机制提供形态学依据。方法 20只雄性Wistar大鼠被随机分为2组,12只行袖状胃切除术做为手术组;8只行胃干预组做为假手术组,术前、术后第1天及术后1月内每周,而后每两周进行体重测量,术后12周处死大鼠后制作前胃石蜡切片,组织学方法观察其形态变化。结果手术组大鼠存活10只,成活率83%。假手术组大鼠在术后增重明显,手术组大鼠体重在术后2周恢复术前水平,体重增长明显但较假手术组慢（P〈0.05）。手术组大鼠术后前胃有代偿性扩张,光镜下可见黏膜及黏膜肌层厚度变薄,黏膜皱襞变浅。结论袖状胃切除术大鼠模型中对前胃的形态学观察可能有利于进一步阐明该术式大鼠模型的减重和体重反弹机制。     

Feed intake, forestomach fluid volume, dilution rate and mean retention of fluid in the forestomach during water deprivation and rehydration in camels (Camelus sp.)

Camels were deprived of water for 11 days. Before and during water deprivation and during rehydration changes in body weight, feed and water intake were measured. Using the liquid marker Cr-EDTA forestomach fluid volume, mean fluid retention and fluid dilution in the forestomach were estimated. At the eleventh day of water deprivation hay intake had decreased to only 9.6% of controls, dilution rates had decreased to 31%, mean retention time of fluid in the forestomach had increased to 189%. At the end of dehydration flow of saliva of 2 l/h mainly contributed to the still rather high dilution rates. Thereby buffering capacity and flow of fluid into the forestomach for microbial digestion as well as the outflow from the forestomach were maintained. At the beginning of rehydration camels drank 97 l within a few minutes, and animals thereby replaced all the water lost. Following this first huge water intake water is rapidly absorbed from the forestomach, and forestomach volume decreased again to dehydration values. At the third day of rehydration control values were reached again. Although feed intake decreased dramatically during water deprivation, functions of the forestomach can be maintained sufficiently mainly due to saliva inflow. This explains the mostly rapid recovery of camels when water is available again.     

Possible involvement of NO-mediated oxidative stress in induction of rat forestomach damage and cell proliferation by combined treatment with catechol and sodium nitrite

To clarify the mechanisms underlying forestomach carcinogenesis in rats by co-treatment with catechol and sodium nitrite (NaNO2), we investigated the involvement of oxidative stress resulting from reaction of the two compounds. Since generation of semiquinone radical, hydroxyl radical (*OH), and peroxynitrite (ONOO-) arose through the reaction of catechol with NO, we proposed that superoxide resulting from catechol oxidation reacted with excess NO, consequently yielding *OH via ONOO-. Male F344 rats were co-treated with 0.2% catechol in the diet and 0.8% NaNO2 in the drinking water for 2 weeks. Prior to occurrence of histological evidence indicating epithelial injury and hyperplasia, 8-hydroxydeoxyguanosine levels in forestomach epithelium significantly increased from 12 h together with appearance of immunohistochemically nitrotyrosine-positive epithelial cells. There were no remarkable changes in rats given each chemical alone. We conclude that oxidative stress due to NO plays an important role in induction of forestomach epithelial damage, cell proliferation, and thus presumably forestomach carcinogenesis.     

The intermediate filament system of the keratinizing mouse forestomach epithelium: Coexpression of keratins of internal squamous epithelia and of epidermal keratins in differentiating cells

Summary The internal epithelium of mouse forestomach represents a fully keratinized tissue that has many morphological aspects in common with the integumental epidermis. In the present study we have, therefore, analyzed keratin expression in the total epithelium, in subfractions of basal cells and in living and dead suprabasal cells that were obtained by Percoll density gradient centrifugation of trypsin-dissociated forestomach keratinocytes. The keratin analysis revealed that basal forestomach keratinocytes synthesize the same keratin types as basal epidermal cells (60 000, 52 000 and 47 000 daltons), whereas differentiating cells contain both the epidermal suprabasal keratin pair (67 000 and 59 000 daltons) and the suprabasal keratin pair characteristic for other internal squamous epithelia (57 000 and 47 000 daltons). Indirect immunofluorescence using an antibody recognizing the members of the epidermal-type suprabasal keratin pair and in-situ-hybridization experiments using specific cDNA probes for the members of the internal-type keratin pair showed that the two keratin pairs are uniformly coexpressed in living suprabasal forestomach keratinocytes. Furthermore, it could be shown that distinct cells in the basal cell layer acquire the ability to express both the 67 000/59 000 dalton and the 57 000/47 000 dalton keratin pair and that some basal cells apparently lose the ability to synthesize mRNAs for basal keratins.     

Studies on the in vitro cultivation of ciliate protozoa from the kangaroo forestomach

The methods used for culturing rumen protozoa were found to be unsatisfactory for growth of ciliate protozoa from the kangaroo forestomach. Based on published measurements of physical parameters in the marsupial forestomach, several modifications were incorporated into the procedure, i.e., an increase in % hydrogen in the gas phase, adjustment of initial pH of the medium to 6.9–7.0 range, feed only forage as a substrate and incubate at a lower temperature (33–36 °C). Only incubation at the lower temperature increased survival time of the kangaroo protozoa. Two species of Bitricha were still viable after 28 d in culture. Cultures had to be terminated at that time. One of the species differed considerably in size and shape from previously described species and based on 18S rRNA data, may represent a new species of Bitricha. The second species, present in low numbers was identified as Bitricha oblata. In a separate trial, Macropodinium yalanbense survived for 11 d, at which time these cultures also had to be terminated.