Atmospheric deposition, CO2, and change in the land carbon sink.

Concentrations of atmospheric carbon dioxide (CO2) have continued to increase whereas atmospheric deposition of sulphur and nitrogen has declined in Europe and the USA during recent decades. Using time series of flux observations from 23 forests distributed throughout Europe and the USA, and generalised mixed models, we found that forest-level net ecosystem production and gross primary production have increased by 1% annually from 1995 to 2011. Statistical models indicated that increasing atmospheric CO2 was the most important factor driving the increasing strength of carbon sinks in these forests. We also found that the reduction of sulphur deposition in Europe and the USA lead to higher recovery in ecosystem respiration than in gross primary production, thus limiting the increase of carbon sequestration. By contrast, trends in climate and nitrogen deposition did not significantly contribute to changing carbon fluxes during the studied period. Our findings support the hypothesis of a general CO2-fertilization effect on vegetation growth and suggest that, so far unknown, sulphur deposition plays a significant role in the carbon balance of forests in industrialized regions. Our results show the need to include the effects of changing atmospheric composition, beyond CO2, to assess future dynamics of carbon-climate feedbacks not currently considered in earth system/climate modelling.

Effects of castration and weaning conducted concurrently or consecutively on behaviour, blood traits and performance in beef calves.

The objectives of this study were to evaluate the effects of Burdizzo castration and abrupt weaning on the behaviour, blood traits and performance of beef calves when weaning was conducted concurrently or consecutively to castration. In total, 64 male beef calves aged between 6 and 7 months were assigned to a 2×2 factorial design with the following treatment groups (n=16 animals per treatment): (1) castrated and concurrently weaned in week 0 (CAS-WEA); (2) castrated in week 0 and weaned in week 4 (CAS-CON); (3) bulls weaned in week 0 (BUL-WEA); and (4) bulls weaned in week 4 (BUL-CON). The behaviour of the calves was observed for 3 days following weaning. Blood was collected weekly from weeks 0 to 5 and analysed for the acute-phase protein haptoglobin, and neutrophil and lymphocyte percentages. BW was recorded weekly from weeks 0 to 7. Animals were slaughtered at 17 months and weight, dressing percentage and carcass classifications were recorded. On day 1 after weaning, the number of vocalizations (calls/10 min) was higher in BUL-WEA (7.2) and CAS-WEA (5.4) than in calves of CAS-CON (2.8) and BUL-CON (2.9) groups (P<0.05). From days 1 to 3 vocalizations decreased in all groups. CAS-CON and BUL-CON animals spent 20% lying on day 1 after weaning compared with 40% in CAS-WEA and BUL-WEA calves (P<0.05). The haptoglobin concentration decreased during the first 5 weeks after weaning in all groups independent of the castration, weaning group or its interaction (P>0.05). WEA groups showed an increased average daily gain (ADG) during weeks 0 to 3 and a reduced ADG during 4 to 7 weeks in comparison with CON animals. At slaughter, bulls were about 80 kg heavier than castrates and had a superior dressing percentage and carcass classification (P>0.05). In conclusion, weaning had a greater effect on the number of vocalizations, standing/walking and lying behaviour and ADG compared with Burdizzo castration. In comparison with undertaking the procedures separately, concurrent castration and weaning neither affected behaviour and haematological parameters nor impaired performance. There was no evidence that the concurrent application of both treatments markedly increased the stress response compared with their application at intervals of a few weeks.

Carcass Characteristics and Meat Quality of Swamp Buffaloes (Bubalus bubalis) Fattened at Different Feeding Intensities.

Twenty-four male 1-year old swamp buffaloes (Bubalus bubalis) were randomly allocated to 4 groups. One group grazed on guinea grass (GG) and another on guinea grass and the legume Stylosanthes guianensis (GL). The other two groups were kept in pens and fed freshly cut guinea grass and concentrate at an amount of 1.5% (GC1.5) and 2.0% (GC2.0) of body weight, respectively. The effect of the different feeding intensities on carcass characteristics and meat quality were assessed. The mean body weight at slaughter was 398 (±16) kg. Average daily gain was higher in concentrate-supplemented groups (570 and 540 g/d in GC1.5 and GC2.0, respectively) when compared to GG (316 g/d) and GL (354 g/d) (p<0.01). Likewise, the warm carcass weight was higher in GC1.5 and GC2.0 compared to GG and GL. Dressing percentage was 48.1% and 49.5% in GC1.5 and GC2.0 in comparison to 42.9% and 44.8% observed in GG and GL, respectively. Meat of Longissimus throracis from GC1.5 and GC2.0 was redder in color (p<0.01), while water holding capacity (drip and thawing loss) was improved in pasture-fed groups (p<0.05). Protein and fat content of Longissimus thoracis was higher in animals supplemented with concentrate (p<0.01), as was cholesterol content (p<0.05), whereas PUFA:SFA ratio was higher and n-6/n-3 ratio lower (p<0.01) in pasture-fed buffaloes. Results of the present study showed that the supplementation of pasture with concentrate enhances the growth and carcass characteristics of swamp buffaloes expressed in superior dressing percentage, better muscling, and redder meat with a higher content of protein and fat, whereas animals grazing only on pasture had a more favorable fatty acid profile and water holding capacity. In conclusion, the supplementation of concentrate at a rate of about 1.5% of body weight is recommended to improve the performance and carcass quality of buffaloes.

The risk-based approach to ATMP development - generally accepted by regulators but infrequently used by companies.

Advanced therapy medicinal products (ATMPs) are the cutting edge of drug innovation. ATMPs have different challenges than other drug classes. To accommodate these challenges and facilitate science-driven development, flexibility in the requirements to demonstrate the safety and efficacy of this rapidly evolving drug class is necessary. To create flexibility, the European Union introduced the risk-based approach. This approach provides the possibility of omitting guideline-based studies based on risk analyses. To gain insight into the effect of the risk-based approach on the non-clinical development of ATMPs, two questions are addressed in this paper. Firstly, "Do companies use a risk-based approach for the non-clinical development of ATMPs?" and, secondly, "Does the Committee for Medicinal Products for Human Use (CHMP) of the European Medicines Agency (EMA) accept non-clinical development programs based on the risk-based approach?" Scientific advice letters formulated by the CHMP were analyzed. The risk-based approach was used to justify deviations from the guidelines in the majority (75%) of the cases. The CHMP accepted 40% of the proposals to omit studies and stated that additional data was necessary to make an informed decision for 35% of the proposals. This indicates that the risk-based approach facilitates the science-driven development of ATMPs.

Future water resources for food production in five South Asian river basins and potential for adaptation--a modeling study.

The Indian subcontinent faces a population increase from 1.6 billion in 2000 towards 2 billion around 2050. Therefore, expansion of agricultural area combined with increases in productivity will be necessary to produce the food needed in the future. However, with pressure on water resources already being high, and potential effects of climate change still uncertain, the question rises whether there will be enough water resources available to sustain this production. The objective of this study is to make a spatially explicit quantitative analysis of water requirements and availability for current and future food production in five South Asian basins (Indus, Ganges, Brahmaputra, Godavari and Krishna), in the absence or presence of two different adaptation strategies: an overall improvement in irrigation efficiency, and an increase of reservoir storage capacity. The analysis is performed by using the coupled hydrology and crop production model LPJmL. It is found that the Godavari and Krishna basins will benefit most from an increased storage capacity, whereas in the Ganges and the Indus water scarcity mainly takes place in areas where this additional storage would not provide additional utility. Increasing the irrigation efficiency will be beneficial in all basins, but most in the Indus and Ganges, as it decreases the pressure on groundwater resources and decreases the fraction of food production that would become at risk because of water shortage. A combination of both options seems to be the best strategy in all basins. The large-scale model used in this study is suitable to identify hotspot areas and support the first step in the policy process, but the final design and implementation of adaptation options requires supporting studies at finer scales.

Short communication: milk output in llamas (Lama glama) in relation to energy intake and water turnover measured by an isotope dilution technique.

Despite the fact that llamas have become increasingly popular as companion and farm animals in both Europe and North America, scientific knowledge on their nutrient requirements is scarce. Compared with other livestock species, relatively little is known especially about the nutrient and energy requirements for lactating llamas. Therefore, we aimed to measure milk output in llama dams using an isotope dilution technique and relate it to energy intakes at different stages of lactation. We also validated the dilution technique by measuring total water turnover (TWT) directly and comparing it with values estimated by the isotope dilution technique. Our study involved 5 lactating llama dams and their suckling young. Milk output and TWT were measured at 4 stages of lactation (wk 3, 10, 18, and 26 postpartum). The method involved the application of the stable hydrogen isotope deuterium ((2)H) to the lactating dam. Drinking water intake and TWT decreased significantly with lactation stage, whether estimated by the isotope dilution technique or calculated from drinking water and water ingested from feeds. In contrast, lactation stage had no effect on dry matter intake, metabolizable energy (ME) intake, or the milk water fraction (i.e., the ratio between milk water excreted and TWT). The ratios between TWT measured and TWT estimated (by isotope dilution) did not differ with lactation stage and were close to 100% in all measurement weeks, indicating that the D(2)O dilution technique estimated TWT with high accuracy and only small variations. Calculating the required ME intakes for lactation from milk output data and gross energy content of milk revealed that, with increasing lactation stage, ME requirements per day for lactation decreased but remained constant per kilogram of milk output. Total measured ME intakes at different stages of lactation were similar to calculated ME intakes from published recommendation models for llamas.

[Recommendations for the husbandry and welfare of sheep and goats by the German Small Ruminant Veterinary Association. Part 2]. / Empfehlung für die Haltung von Schafen und Ziegen der Deutschen Gesellschaft für die Krankheiten der kleinen Wiederkäuer, Fachgruppe der DVG. Teil 2.

The second part of the recommendations deals with the healthcare and the regulatory framework for the husbandry of sheep and goats. The suggested concept for healthcare aims to develop an individual health plan for every flock. This health plan focuses not only on the prevention of notifiable diseases, but also on chronic and slow infections as well as on parasite monitoring. The emphasis is on early detection of diseases and prophylaxis. In conjunction with this, the handling of lameness, shearing, animal trade and quarantine as well as cleaning and disinfection in sheep and goat flocks are intensively discussed. There are detailed federal and European legal regulations concerning the transport and the physical well-being of animals. These laws are clearly presented and advice for their practical implementation is provided.

[Infectious causes of lameness in sheep - treatment options in veterinary practice]. / Infektiöse Lahmheiten beim Schaf - Therapiemöglichkeiten in der tierärztlichen Praxis.

Lameness in sheep induced by infectious agents can cause problems regarding animal welfare, diagnosis and treatment. Individual lame animals and lameness caused by systemic disease, but especially flock problems due to infectious causes of lameness, such as foot rot, can for various reasons pose a problem for veterinary treatment. The causes of lameness in sheep are described with a special focus on infectious flock problems, and the different treatment options such as foot trimming, foot baths and local and systemic antibiotic therapy as well as vaccination are discussed based on a review of recent international research. The limited choice of drugs licensed for use in sheep in Germany is highlighted. Treatment, therefore, often requires off-label use or the import of footbathing agents licensed in other European countries. The legal consequences of this lack of nationally licensed veterinary products in dealing with a "minor species" are discussed, with a final call for political solutions that will help improve this unsatisfactory situation.

[Recommendations for the husbandry and welfare of sheep and goats by the German Small Ruminant Veterinary Association. Part 1]. / Empfehlung für die Haltung von Schafen und Ziegen der Deutschen Gesellschaft für die Krankheiten der kleinen Wiederkäuer, Fachgruppe der DVG. Teil 1.

Recommendations for the different forms of sheep and goat husbandry based on the legal regulations are summarized. These are given in particular respect to transhumance, tending, alpine farming, and indoor housing. The requirements for pasture, housing, supply of water and food, lambing, rearing of lambs, and health management are intensively discussed. The general requirements of the extensive as well as of the intensive husbandry of sheep and goats are defined. Examples of species-specific capabilities for adaption, the limits of adaption, and signs of decompensation are provided. Compliance with these recommendations should accord the animals entrusted to our care the "five freedoms (13)": 1. Freedom from hunger and thirst, 2. freedom from discomfort, 3. freedom from pain, injury, or disease, 4. freedom to express normal behaviour, and 5. freedom from fear and distress.

Is the establishment rate and fecundity of Haemonchus contortus related to body or abomasal measurements in sheep?

The relationship among parasitological parameters, abomasal size and body size measurements was investigated in lambs following an experimental infection with Haemonchus contortus. In total, 100 lambs from five different genotypes (German Merino (GM), Texel × GM, Suffolk × GM, German Blackhead Mutton × GM and Ile de France × GM) were experimentally infected with 5000 infective third stage larvae of H. contortus at the time of weaning at 12 weeks of age. Four and six weeks after infection, individual faecal samples were collected for estimation of faecal egg counts (FECs). Furthermore, wither height, shoulder width, heart girth, loin girth and body length were taken at 18 weeks of life. Lambs were slaughtered and necropsied 7 weeks post-infection, and worm counts, abomasal volume and surface area were determined. Positive correlations were found between different body size parameters, body weight and abomasal sizes. FEC and worm counts were not significantly correlated either with body size parameters or with abomasal size. The mean worm burden was higher in GM than in crossbred lambs. There was no significant difference in abomasal size between GM and crossbred lambs. The results suggest that the variations between animals in worm burden following an experimental infection with H. contortus (worm resistance) are not influenced by body size parameters or abomasal sizes. Therefore, other factors, including genetic-based differences in resistance, must cause these findings between and within breeds.

Conditional approval and approval under exceptional circumstances as regulatory instruments for stimulating responsible drug innovation in Europe.

The need for fast drug innovation and the public demand for risk-free drugs creates a dilemma for regulatory authorities: less restrictive procedures involve uncertainties about benefit/risk profiles of new drugs. The European Union has introduced two instruments that regulate early market access: conditional approvals (CAs) and approvals under exceptional circumstances (ECs). We have studied whether these instruments compromise the safety of new drugs and whether they lead to earlier access to innovative drugs. Our study shows that neither of these regulatory pathways accelerates the approval process for innovative drugs. However, the CA pathway shortens the clinical development period. Approvals under ECs are associated with longer clinical development periods, but this regulatory pathway may open up opportunities for specific drugs to be admitted into the market because less comprehensive data are required. Despite the fact that these advanced approvals are based on limited safety databases, there are no special safety issues associated with using these pathways.

Ascaridia galli infection of pullets and intestinal viscosity: consequences for nutrient retention and gut morphology.

1. Pullets were given a control diet or a diet supplemented with a non-starch-polysaccharide hydrolysing enzyme preparation (NSP-enzyme) from weeks 6 to 14 of age to induce differences in the viscosity of the small intestinal ingesta. Half of each feeding group (n = 25) was infected with 250 embryonated Ascaridia galli eggs at an age of 6 weeks. 2. At 14 weeks of age, before the pullets were slaughtered, a balance experiment was conducted, to sample ingesta for viscosity measurements. Also, tissue samples of jejunum and ileum were taken for morphometrical and histopathological studies. 3. An infection of pullets with A. galli reduced the viscosity of the jejunal ingesta at high initial levels of viscosity after feeding the NSP-enzyme unsupplemented diet. 4. The faecal A. galli egg output by the pullets expressed as eggs per g excrement (EpG) was significantly reduced in infected pullets given the NSP-enzyme supplemented diet. Also, the number and length of worms was less in these pullets. 5. The tunica muscularis of the jejunum was significantly thickened, this effect being more pronounced at a low intestinal viscosity. 6. NSP-enzyme addition resulted in an increased length of jejunal villus and was paralleled by a decrease in jejunal viscosity. 7. Histopathology of jejunal and ileal sections revealed no pathological alterations. 8. The apparent retention of organic matter was increased after enzyme addition while parasite infection exerted no effect.

Genetic resistance to Heterakis gallinarum in two chicken layer lines following a single dose infection.

Groups of White Leghorn and New Hampshire male and female chicks were reared under helminth-free conditions and divided into two groups at an age of 8 weeks. One group was artificially infected with 100 embryonated Heterakis gallinarum eggs, while the other group was kept as uninfected control. After 8 weeks all animals were harvested and worm burden was established. Maximum of 60 worms per bird randomly selected were measured for length. Body weights were recorded at 8 and 16 weeks of age. Total worm burden was significantly (P<0.0001) higher in infected males than in infected females. Furthermore, White Leghorns showed a significantly (P=0.011) higher number of worms when compared with New Hampshire animals. Average worm lengths were not significantly different between the breeds (P=0.155) and sexes (P=0.062). Correlations between worm burden and worm length was 0.08 (P=0.140). The daily weight gains between 8 and 16 weeks of age of the infected animals were significantly (P=0.038) lower when compared with the control group. There was no significant correlation between daily weight gain and worm burden. The estimated heritabilities for logarithm (ln) worm burden were 0.41 (S.E.+/-0.086) in White Leghorn and 0.31 (S.E.+/-0.126) in New Hampshire, respectively. Based on the estimated heritabilities it is possible to select for H. gallinarum resistance in both genetically distinct chicken breeds. This approach may be of importance for chickens kept in floor husbandry systems.

The role of preparation technique, culture media and incubation time for embryonation of Heterakis gallinarum eggs.

The importance of preparation technique, culture media and incubation time in the embryonation of the infective egg stages of the intestinal nematode parasite Heterakis gallinarum was studied. Mature H. gallinarum worms were isolated from the caeca of infected chickens and separated by sex. In a first experiment intact female worms were kept for the development of their eggs in four different media (0.5% formalin, 2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) and incubated under constant temperature (20-22 degrees C) for 2, 4, 6 or 8 weeks. Afterwards the body of the worms were ruptured and the numbers of unembryonated and embryonated eggs were determined using a McMaster egg counting chamber, and the percentage of embryonated eggs was calculated. After 8 weeks of incubation in 0.5% formalin, 0.1 N sulphuric acid or 0.1% potassium dichromate 27.6%, 26.7% and 29.4% of the eggs, respectively, embryonated into third stage larvae (p > 0.05). In contrast, incubation in 2% formalin resulted in an embryonation of 18.6% only (p < 0.05). In a second experiment H. gallinarum eggs were directly harvested from worm uteri and cultivated afterwards in different media (2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) at 20 to 22 degrees C for 6 weeks. An incubation of isolated eggs in 2.0% formalin or 0.1% potassium dichromate during 6 weeks resulted in a significantly higher percentage of embryonation in comparison to the incubation of intact worms (first experiment). The results suggest that preparation technique, media and time of incubation has an essential influence on the development rate of H. gallinarum eggs.

Effects of weaning lambs in two stages or by abrupt separation on their behavior and growth rate.

A study was conducted to evaluate effects of age at weaning in combination with different weaning procedures on 2 breeds of lambs (Merinoland, Rhoenschaf). Lambs were either weaned at 8 or 16 wk of age in 2 stages or with the traditional method of weaning by abrupt separation. In the 2-stage treatment, lambs were prevented from nursing their dam for 1 wk (stage 1) before their separation (stage 2). Control animals were nursed by their dams until they were separated. Lamb BW and behavior (vocalization, agitation) were recorded before and after separation. After separation, lambs weaned at 8 wk of age had greater (P = 0.004) ADG compared with lambs weaned at 16 wk of age. Control lambs had greater (P < 0.001) agitation scores [1 = no agitation (normal behavior; i.e., feeding, resting, lying, standing, or play behavior) to 3 = high agitation (continuously moving, restlessness, or vocalization)] irrespective of weaning age and breed. On the day of separation, 2-stage lambs had scores from 1.17 to 1.35, whereas control lambs were scored from 1.70 to 1.79. After separation, lambs weaned in 2 stages vocalized up to 98.2% less (P = 0.001) than control animals. Furthermore, bleats were greater for Rhoenschaf lambs and at a weaning age of 8 wk (P < 0.05). Differences between treatments were greatest on the day of separation. Vocalization decreased continuously within the first 3 d of weaning to zero. Lambs weaned in 2 stages were less distressed than lambs weaned by the traditional method of abrupt separation based on behavioral data, but ADG until 12 and 16 wk of age did not differ (P > 0.05) for either treatment in this study.

Measurement of milk intake in suckling llamas (Lama glama) using deuterium oxide dilution.

The objective of the study was to estimate daily milk intake in llama crias and relate nutrient intakes at peak lactation to growth data. Milk intake in 11 suckling llamas was estimated from water kinetics using deuterium oxide (D2O) at d 17, 66, and 128 postpartum. Daily milk intakes averaged 2.6, 2.3, and 2.0 kg at 17, 66, and 128 d postpartum, respectively. Milk intake decreased with age when expressed as daily amount, percentage of body weight (BW), or per kilogram of metabolic size, but the influence of age was eliminated when expressed per gram of daily gain. Because llamas only have one young per parturition, milk intake was equivalent to the daily milk output of the dam, which ranged from 27.6 to 96.9 g/kg of maternal BW(0.75). Compared with different ruminant species, milk production in llamas appears to lie between wild and domestic ruminants used for meat production. Nutrients (dry matter, fat, protein, and lactose) and energy intakes from the milk calculated by combining milk intake and milk composition data decreased with age when expressed as daily amount or per 100 g of BW, but when expressed per gram of daily gain, no clear trend was observed. Maintenance requirement for suckling llamas at peak lactation (17 d postpartum) was 312 kJ of ME/kg of BW(0.83). Combined with milk composition data, the present milk intake estimations at different stages of the lactation can be used to establish recommendations for nutrient and energy requirements of suckling llamas.

Respiration as the main determinant of carbon balance in European forests.

Carbon exchange between the terrestrial biosphere and the atmosphere is one of the key processes that need to be assessed in the context of the Kyoto Protocol. Several studies suggest that the terrestrial biosphere is gaining carbon, but these estimates are obtained primarily by indirect methods, and the factors that control terrestrial carbon exchange, its magnitude and primary locations, are under debate. Here we present data of net ecosystem carbon exchange, collected between 1996 and 1998 from 15 European forests, which confirm that many European forest ecosystems act as carbon sinks. The annual carbon balances range from an uptake of 6.6 tonnes of carbon per hectare per year to a release of nearly 1 t C ha(-1) yr(-1), with a large variability between forests. The data show a significant increase of carbon uptake with decreasing latitude, whereas the gross primary production seems to be largely independent of latitude. Our observations indicate that, in general, ecosystem respiration determines net ecosystem carbon exchange. Also, for an accurate assessment of the carbon balance in a particular forest ecosystem, remote sensing of the normalized difference vegetation index or estimates based on forest inventories may not be sufficient.

A network of heterogeneous hydrogen bonds in GNRA tetraloops.

RNA hairpin loops containing a GNRA consensus sequence are the most frequently occurring hairpins in a variety of prokaryotic and eukaryotic RNAs. These tetraloops play important functional roles in RNA folding, in RNA-RNA tertiary interactions and as protein binding sites. Homo and heteronuclear NMR spectroscopy have been used to determine the structures of the most abundant members of the GNRA tetraloop family: the GAGA, GCAA and GAAA loops closed by a C-G base pair. Analysis of the structures of these three hairpin loops reveals a network of heterogeneous hydrogen bonds. The loops contain a G-A base pair, a G base-phosphate hydrogen bond and several 2' OH-base hydrogen bonds. These intramolecular interactions and the extensive base stacking in the loop help explain the high thermodynamic stability and give insight into the diverse biological roles of the GNRA RNA hairpins.

Monoclonal antibody MON-114: detection of a marker for neuroendocrine differentiation in human lung cancer.

Mouse monoclonal antibody MON-114 was generated upon immunization with a human small cell lung carcinoma cell line GLC-19. Immunohistochemical analysis of normal tissues with MON-114 showed staining of the adrenal gland, brain and peripheral nerves. With respect to human lung carcinomas, 7 out of 8 small cell lung carcinomas were positively stained as well as 5 out of 5 carcinoid tumors, whereas only 4 out of 31 squamous cell carcinomas and 3 out of 19 adenocarcinomas were weakly stained. Furthermore, 1 large cell carcinoma was negative for MON-114 staining. Apparently, MON-114 stains cells of neuroendocrine differentiation.