Identification and functional expression of the mitochondrial pyruvate carrier

The transport of pyruvate, the end product of glycolysis, into mitochondria is an essential process that provides the organelle with a major oxidative fuel. Although the existence of a specific mitochondrial pyruvate carrier (MPC) has been anticipated, its molecular identity remained unknown. We report that MPC is a heterocomplex formed by two members of a family of previously uncharacterized membrane proteins that are conserved from yeast to mammals. Members of the MPC family were found in the inner mitochondrial membrane, and yeast mutants lacking MPC proteins showed severe defects in mitochondrial pyruvate uptake. Coexpression of mouse MPC1 and MPC2 in Lactococcus lactis promoted transport of pyruvate across the membrane. These observations firmly establish these proteins as essential components of the MPC.     

Influence of floor surface and access to pasture on claw health in dairy cows kept in cubicle housing systems

In this study, the effects on the claw health of dairy cows of three different floor types and access to pasture were investigated on 35 farms. The farms were fitted with a given floor type in the indoor walking area of a cubicle housing system: a solid rubber, mastic asphalt or slatted concrete floor. Because we chose farms on which the given floor type was in good condition, the data presented show what can be achieved on these types of floors under ideal circumstances. Cows on half of the farms per floor type had access to pasture during the grazing period. Each farm was visited three times at approx. 6-month intervals at the end of the winter indoor-housing period and at the end of the summer period, i.e. after the period with access to pasture on half of the farms. During each visit, the claw health of the same 10 cows per farm was assessed on the occasion of routine claw trimming. The proportion of cows with haemorrhages increased from mastic asphalt to rubber and slatted concrete floors. A lower proportion of cows kept on mastic asphalt was affected by white-line fissures and needed intermittent claw-trimming, an indicator for lameness. Cows housed in cubicle systems with slatted concrete floors were at the lowest risk of having heel-horn erosions. Access to pasture was associated with a lower incidence of slight white-line fissures and dermatitis digitalis. A higher proportion of cows with sole haemorrhages and sole ulcers were found on all floor types at the end of the summer period than at the end of the winter indoor-housing period. Floor type did not influence the presence of sole ulcers and deep white-line fissures. In conclusion, the effect of floor type on claw health was slight, and none of the investigated floor types was clearly superior to the others. Access to pasture was not effective in reducing the presence of most types of claw lesions associated with the floor type used in the indoor walking area.     

Development of an express method for measuring soil nitrate,phosphate, potassium,and pH for future in-field application

Background

In practical farming, there is often a need for short-term availability of information on the soil nutrient status.

Aims

To develop a new express method for the extraction of major plant-available nutrients and measurement of soil nutrients. In future, this method shall serve for in-field measurements of soil samples with an ion-sensitive field-effect transistor (ISFET).

Methods

Various extraction conditions such as type of extractant, soil-to-solution ratio, time, and intensity were investigated on a broad selection of dried soil samples in the laboratory. Based on 83 field-moist soil samples with varying clay contents, these conditions were compared to standard laboratory methods.

Results

With increasing extraction time, the nutrient concentrations increased. When the soil-to-solution ratio was reduced, a greater share of nutrients was extracted, independent of soil type. H₂O and 0.01 M CaCl₂ and standard calcium-acetate-lactate (CAL) solution proved to be too weak in the short period to reach the ISFET sensor measurement range. Higher concentrated CAL solutions performed much better. Finally, a 5-min CaCl₂ extraction followed by the removal of an aliquot for the determination of soil pH and NO₃⁻ was found to be effective. The remaining solution was then mixed with 0.20 M CAL solution for the analysis of H₂PO₄⁻ and K⁺ at 10 min of extra extraction time. This extraction method showed very good correlations with the values based on the German laboratory reference methods for pH (R² = 0.91) and for nitrate (R² = 0.95). For phosphorus and potassium, we obtained an R² of 0.70 and 0.81, respectively, for all soils. When soils were grouped according to clay content higher correlations were found.

Conclusions

A new express method based on a wet-chemical approach with a soil preparation procedure was successfully developed and validated. This seems to be a valuable basis for future in-field measurements via ISFET.     

The prevalence and clinical relevance of hyperkalaemia in calves with neonatal diarrhoea

One hundred and twenty-four calves with neonatal diarrhoea were investigated in order to assess the prevalence of hyperkalaemia and the associated clinical signs. Hyperkalaemia (potassium concentration >5.8 mmol/L) was recognized in 42 (34%) calves and was more closely associated with dehydration than with decreases in base excess or venous blood pH. In 75 calves with normal blood concentrations of D-lactate (i.e. ?3.96 mmol/L), K concentrations were moderately correlated with base excess values (r = ?0.48, P < 0.001). In contrast, no significant correlation was observed in 49 calves with elevated D-lactate. Only three hyperkalaemic calves had bradycardia and a weak positive correlation was found between heart rate and K concentrations (r = 0.22, P = 0.014). Ten of the 124 calves had cardiac arrhythmia and of these seven had hyperkalaemia indicating that cardiac arrhythmia had a low sensitivity (17%) but a high specificity (96%) as a predictor of hyperkalaemia.In a subset of 34 calves with base excess values ??5 mmol/L and D-lactate concentrations <5 mmol/L (of which 22 had hyperkalaemia), changes in posture/ability to stand could be mainly explained by elevations of K concentrations (P < 0.001) and to a lesser extent by increases in L-lactate concentrations (P = 0.024). Skeletal muscle weakness due to hyperkalaemia alongside hypovolaemia may produce a clinical picture that is similar to that in calves with marked D-lactic acidosis. However, since reductions in the strength of the palpebral reflex are closely correlated with D-lactate concentrations, a prompt palpebral reflex can assist the clinical prediction of hyperkalaemia in calves presenting with a distinct impairment in their ability to stand (specificity 99%, sensitivity 29%).     

Attenuation of virulence of Lawsonia intracellularis after in vitro passages and its effects on the experimental reproduction of porcine proliferative enteropathy

Non-pathogenic Lawsonia intracellularis variants have been obtained through multiple passages in cell culture but there is no information regarding the number of passages necessary to attenuate a pathogenic isolate. The present study evaluated the susceptibility of pigs to L. intracellularis after 10, 20 and 40 passages in vitro. Three groups (six animals/group) were inoculated with pure culture of L. intracellularis on passage 10, 20 or 40 and one group with placebo. The animals were monitored for clinical signs, fecal shedding and serological IgG response during 28 days post-inoculation. Gross and histologic lesions and the level of infection based on the amount of L. intracellularis-specific antigen in the intestinal mucosa identified by immunohistochemistry were evaluated in two animals from each group on days 14, 21 and 28. Animals inoculated with passages 10 and 20 demonstrated proliferative lesions typical of porcine proliferative enteropathy associated with the presence of Lawsonia-specific antigen in the intestinal mucosa. Passage 40-inoculated pigs did not show proliferative lesions or presence of Lawsonia antigen at any time point throughout the study. Similar patterns of the fecal shedding were observed in passage 10 and 20-infected pigs but those infected with passage 40 shed for a short period. Serological IgG responses in passage 10 and 20-inoculated pigs were detected from day 14 post-infection but not at all in passage 40-inoculated animals. These results demonstrate attenuation of the virulence properties of L. intracellularis between 20 and 40 cell passages in vitro. This information will be valuable for design of future experimental models and for studying the mechanisms involved in the attenuation of L. intracellularis virulence.     

The base saturation in acidified Swiss forest soils on calcareous and noncalcareous parent material. A pH–base saturation anomaly

Long‐term soil acidification leads to lower pH values and to a concomitant decrease in base saturation (BS). The relationship between pH and base saturation (BS) in acidified forest soils can be disturbed by processes such as nutrient cycling by vegetation, temporary saturation by ground water that comes into contact with calcareous material, or by upward diffusion of base cations from deeper horizons. This paper examines the relationship between pH and BS in Swiss forest soils developed from calcareous and noncalcareous parent material and identifies some of the factors that can affect the BS in the decalcified parts of soils derived from calcareous parent material. The decalcified zone in the latter soils has a higher BS on average compared to soils from noncalcareous parent material, but their pH values are identical. In the pH range 4.0–4.5, the difference in BS may vary by a factor of three. The mean BS in the decalcified zone tends to decrease with increasing depth of the calcareous layer. The water regime also affects the BS in soils on calcareous parent material. In soils temporarily saturated by groundwater (gleysols), the BS in the decalcified zone is always high (85%–100%) because of the continuous contact between the soil water and the calcareous parent material. In addition, the inhibited drainage impedes the depletion of base cations in these soils. In contrast, soils that are temporarily saturated by rainwater are depleted in base cations due to the alternating wetting‐and‐drying regime and the associated leaching of dissolved ions. In such soils, the depletion of base cations is strongly related to the extent of hydromorphy. Stagnogleyic soils, with the longest period of water saturation, have the highest depletion levels. We conclude that in such soils, the diffusion of base cations from deeper zones is strongly compensated by leaching from the very acidic soil horizons. The pH–base saturation anomaly has consequences for some of the methods used to calculate the critical loads of acidity for forest soils in Switzerland, with many soils being less sensitive than previously reported.     

Flavonoids of white lupin roots participate in phosphorus mobilization from soil

The impact of flavonoids released by phosphorus-deficient white lupin roots on inorganic P and soil microorganisms is largely unknown. We report that flavonoids isolated from white lupin roots mobilized inorganic phosphorus and decreased soil microbial respiration, citrate mineralization, and soil phosphohydrolase activities, but did not reduce the soil ATP content. The results suggest that white lupin's release of flavonoids into the rhizosphere plays a significant role in its efficient P-acquisition strategy by solubilizing Fe-bound P and by limiting the microbial mineralization of citrate.     

Heat,wheat and CO2: The relevance of timing and the mode of temperature stress on biomass and yield

Atmospheric CO₂ enrichment affects C3 crops both directly via increased carbon gain and improved water use efficiency and indirectly via higher temperatures and more frequent climatic extremes. Here we investigated the response of spring wheat (Triticum aestivum L. cv. Triso) to CO₂ enrichment (550 vs. 380 µmol/mol) and heat, applied as a constant +4°C increase or a typical heat wave either before or after anthesis, or as two typical heat waves before and after anthesis. We applied a climate chamber approach closely mimicking ambient conditions. CO₂ enrichment increased above‐ground biomass and yield by c. 7 and 10%, but was not able to compensate for adverse heat stress effects, neither before nor after anthesis, with few exceptions only. Yield depression due to heat stress was most severe when two heat waves were applied (?19%). This adverse effect was, however, compensated by CO₂ enrichment. Applying heat stress before or after anthesis did not exert different effects on yield for both +4°C warming and heat wave application. However, +4°C depressed yield more than a heat wave at ambient CO₂, but not so at elevated CO₂. Thus, the interactive effects were complex and prediction of future wheat yield under CO₂ enrichment and climate extremes deserves more attention.     

Agronomic performance of winter wheat grown under highly divergent soil moisture conditions in rainfed and water‐managed environments

Even in the temperate climates of Europe, increasing early season drought and rising air temperature are presenting new challenges to farmers and wheat breeders. Sixteen winter wheat (Triticum aestivum L.) genotypes consisting of three hybrids, six line cultivars and two breeding lines from Germany as well as five line cultivars from France, Austria, Slovakia, Hungary and the Ukraine (referred to as “exotic” lines) have been included in this study. The genetic materials were evaluated over three growing seasons under a range of soil moisture regimes at the three North German sites Braunschweig (irrigated and drought‐stressed), Warmse (rainfed) and Söllingen (rainfed). The average grain yields in the twelve growth environments (water regime × season combinations) ranged from 6.1 to 13.5 t ha^?1. The exotic lines showed little evidence of specific phenological adaptation to drought although they are frequently faced with water scarcity in their countries of origin. The hybrids and German lines exhibited higher regression coefficients (b_i) to environmental means than the exotic lines, indicating particular adaptation to favourable growing conditions. The phenotypical correlations of grain yield between the various environments were high, ranging for instance from 0.6 to 0.8 for the irrigated and drought‐stressed environments at Braunschweig. It is thus expected that in the foreseeable future continued selection aiming at high yield potential will suffice as a means to counter the expected increase in droughts.