Experimental data show that a significant improvement of growth rate and feed conversion rate of weanling pigs can be achieved by the inclusion of organic acids in the diet. These ergotropic effects have been mainly observed with formic, lactic, sorbic, fumaric, citric and malic acid as well as with different salts of formic acid. The lowering of dietary pH alone, with inorganic acids (o-phosphoric acid, HC1), however failed to show any nutritive efficacy. Studies on the mode of action of organic acids indicate that they cause a higher protein and energy digestibility and retention, an alteration of bacterial populations and metabolites in the gastrointestinal tract and possibly an effect on metabolism. It seems likely that the antimicrobial properties of organic acids and of their salts are of great importance for their beneficial effects in weanling pigs.

This investigation was designed to estimate the Co requirement of growing cattle on the basis of plasma and liver levels of vitamin B12 and folate, plasma levels of homocysteine and methylmalonic acid (MMA) and haematological variables. For this purpose thirty-four male intact cattle of the German Simmental breed (236 kg) were assigned randomly to ten groups and were fed corn silage-based diets which contained 70, 90, 109, 147, 184, 257, 327, 421, 589 or 689 μg Co/kg DM for 40 weeks. One-slope broken-line model analysis and a quadratic model with plateau were used to estimate the Co requirement. The broken-line model estimated the dietary Co requirement of growing cattle to be 257 (SE 29) ΜG/KG DIETARY DM BASED ON PLASMA VITAMIN B12 AS RESPONSE CRITERION. THE DIETARY CO LEVELS NEEDED TO MAXIMISE THE LIVER VITAMIN B12 AND LIVER FOLATE WERE 236 (se 8) and 190 (se 8) μg/kg dietary DM respectively. Plasma folate did not show any response to the different Co levels. The dietary Co was inversely correlated with the plasma concentrations of homocysteine and MMA. Estimates of the dietary Co concentration required to minimise homocysteine were 161 (se 10) μg/kg DM. When MMA was used as response criterion, the linear model yielded a Co requirement of 124 (se 3) μg/kg dietary DM. The quadratic model did not provide a better closeness of regression fit and yielded similar requirements to the linear model. Haemoglobin concentration and haematocrit tended to have a slight response to increasing dietary Co and were only decreased in cattle on diets containing less than 100 μg Co/kg DM. On the basis of the present data, recommended levels of dietary Co for normal folate metabolism and minimum homocysteine and MMA levels can be set to be 150–200 μg/kg DM; for maximum vitamin B12 levels, the desired Co content in the diet seems to be 250 μg/kg DM.

Dietary zinc deficiency in rats causes increased osmotic fragility of their erythrocytes. In this study, the influence of supplementary antioxidants (vitamin C, vitamin E or beta-carotene) on osmotic fragility, oxidative damage and components of the primary defense system of erythrocytes of zinc-deficient rats was investigated. Indicators of hemolysis in vivo were also examined. Five groups of 12 male rats were force-fed a zinc-adequate diet (control rats), a zinc-deficient diet or a zinc-deficient diet enriched with vitamin C, vitamin E or beta-carotene. Compared with the control rats, the rats fed the zinc-deficient diet without supplementary antioxidants had greater red blood cell osmotic fragility, higher concentrations of thiobarbituric acid-reactive substances and alanine, higher glutathione S-transferase activity, lower concentration of glutathione and activity of glutathione peroxidase as well as lower activity of superoxide dismutase in plasma (P < 0.05). Supplementation with antioxidants generally improved osmotic fragility in zinc-deficient rats without influencing zinc concentration or alkaline phosphatase activity in plasma, indicators of zinc status. At some of the hypotonic saline concentrations tested, vitamin C and beta-carotene significantly affected osmotic fragility. The zinc-deficient rats fed a diet without supplementary antioxidants had significantly higher concentrations of alanine in erythrocytes than the zinc-deficient rats supplemented with vitamin C, vitamin E or beta-carotene and had significantly higher levels of thiobarbituric acid-reactive substances in erythrocytes than the rats supplemented with beta-carotene. There was no indication of hemolysis in vivo in rats fed zinc-deficient diets. The results show that supplementary antioxidants decrease osmotic fragility and oxidative damage of erythrocytes in zinc-deficient rats.

Introduction  In studies on the quantitative mineral metabolism the separation of total faecal mineral excretion into the fraction of dietary and of endogenous origin is often a methodical barrier. Both components cannot be distinguished by standard chemical procedures. But their separation is essential to the quantification of the mineral flux from the diet into the organism and back from tissues into the faecal excretion as it is necessary for example to quantify the bioavailability of dietary mineral sources (K irchgessner et al. 1993). For this purpose, the use of isotopes is an appropriate means. During the last decades, several techniques employing radioactive or stable isotopes have been developed, e.g. the ‘comparative balance method’, the ‘dual tracer’ or ‘double isotope’ technique, methods based on computerized ‘compartment analysis’ and the ‘isotope‐dilution technique’ (e.g. A ubert et al. 1963; T hompson 1965; B elshaw et al. 1974; G ibson et al. 1988). Among these methods, the isotope‐dilution technique, in particular, comprises direct and quantitative measurements of mineral fluxes and provides robust estimates of endogenous faecal excretions as has been shown for a series of macrominerals and trace elements (W eigand and K irchgessner 1976a,b; W eigand et al. 1986a,b; K reuzer and K irchgessner 1991; R euber et al. 1993; K irchgessner et al. 1994; W indisch and K irchgessner 1994; W indisch et al. 1997; G abler et al. 1997). For iodine however, there is no appropriate isotope method available. Therefore, the present experiment was designed to establish the isotope‐dilution technique for iodine. The isotope‐dilution technique is based on a single parenteral injection or a long‐term oral administration of the tracer (W indisch and K irchgessner 1994). After the labelling procedure, all tracer that appears in the faeces is of endogenous origin. The calculation from the quantity of tracer recovered in the faeces to the total amount of endogenous faecal excretion of the respective element is performed by the use of a reference tissue from which the endogenous excretion originates or which is at least in very close physiological relationship to the endogenous excretion. Using 125I as tracer the total amount of endogenous faecal iodine is calculated as follows: Endogenous faecal iodine (ng/day) = Afaeces/SAreference tissueAfaeces = 125I activity in the faeces (Bq/day)SAreference tissue = specific 125I activity of the reference tissue (Bq 125I per ng of total iodine)True absorption of dietary iodine (ng/day) = Iodine intake – iodine in faeces (total – endogenous)In total, the present methodological study had to focus on two major aspects. Since the administered tracer needs time to reach steady‐state kinetics within the excretory pool of iodine it was to be clarified at first, from which day after a single 125I administration does the faecal 125I excretion correctly represent the total endogenous excretion quantitatively. ...

Tight Junctions (TJ) regulate paracellular permeability of tissue barriers. Claudins (Cld) form the backbone of TJ-strands. Pore-forming claudins determine the permeability for ions, whereas that for solutes and macromolecules is assumed to be crucially restricted by the strand morphology (i.e., density, branching and continuity). To investigate determinants of the morphology of TJ-strands we established a novel approach using localization microscopy.TJ-strands were reconstituted by stable transfection of HEK293 cells with the barrier-forming Cld3 or Cld5. Strands were investigated at cell-cell contacts by Spectral Position Determination Microscopy (SPDM), a method of localization microscopy using standard fluorophores. Extended TJ-networks of Cld3-YFP and Cld5-YFP were observed. For each network, 200,000 to 1,100,000 individual molecules were detected with a mean localization accuracy of ∼20 nm, yielding a mean structural resolution of ∼50 nm. Compared to conventional fluorescence microscopy, this strongly improved the visualization of strand networks and enabled quantitative morphometric analysis. Two populations of elliptic meshes (mean diameter <100 nm and 300–600 nm, respectively) were revealed. For Cld5 the two populations were more separated than for Cld3. Discrimination of non-polymeric molecules and molecules within polymeric strands was achieved. For both subtypes of claudins the mean density of detected molecules was similar and estimated to be ∼24 times higher within the strands than outside the strands.The morphometry and single molecule information provided advances the mechanistic analysis of paracellular barriers. Applying this novel method to different TJ-proteins is expected to significantly improve the understanding of TJ on the molecular level.