Alternative methodologies - stretching the in vitro box

Current gas-based in vitro evaluation systems are extremely powerful research techniques. However they have the potential to generate a great deal more than simple fermentation dynamics. Details from four experiments are presented in which adaptation, and novel application, of an in vitro system allowed widely differing objectives to be examined. In the first two studies, complement methodologies were utilised. In such assays, an activity or outcome is inferred through the occurrence of a secondary event rather than by direct observation. Using an N-deficient incubation medium, the increase in starch fermentation, when supplemented with individual amino acids (i.e., known level of N) relative to that of urea (i.e., known quantity and N availability), provided an estimate of their microbial utilisation. Due to the low level of response observed with some arnino acids (notably methionine and lysine), it was concluded, that they may not need to be offered in a rumen-inert form to escape rumen microbial degradation. In another experiment, the extent to which degradation of plant cell wall components was inhibited by lipid supplementation was evaluated using fermentation gas release profiles of washed hay. The different responses due to lipid source and level of inclusion suggested that the degree of rumen protection required to ameliorate this depression was supplement dependent. That in vitro inocula differ in their microbial composition is of little interest per se, as long as the outcome is the same (i.e., that similar substrates are degraded at comparable rates and end-product release is equivalent). However where a microbial population is deficient in a particular activity, increasing the level of inoculation will have no benefit. Estimates of hydrolytic activity were obtained by examining fermentation kinetics of specific substrates. A number of studies identified a fundamental difference between rumen fluid and faecal inocula, with the latter having a lower fibrolytic activity, which could not be completely attributed to microbial numbers. The majority of forage maize is offered as an ensiled feed, however most of the information on which decisions such as choice of variety, crop management and harvesting date are made is based on fresh crop measurements. As such, an attempt was made to estimate ensiled maize quality from an in vitro analysis of the fresh crop. Fermentation profiles and chemical analysis confirmed changes in crop composition over the growing season, and loss of labile carbohydrates during ensiling. In addition, examination of degradation residues allowed metabolizable energy (ME) contents to be estimated. Due to difficulties associated with starch analysis, the observation that this parameter could be predicted by difference (together with an assumed degradability), allowed an estimate of ensiled maize ME to be developed from fresh material. In addition, the contribution of the main carbohydrates towards ME showed the importance of delaying harvest until maximum starch content has been achieved. (c) 2005 Elsevier B.V. All rights reserved.

Logit models for identifying the factors that influence the uptake of new 'no-tillage' technologies by farmers in the rice-wheat and the cotton-wheat farming systems of Pakistan's Punjab

In the 'rice-wheat' and the 'cotton-wheat' farming systems of Pakistan's Punjab, late planting of wheat is a perennial problem due to often delayed harvesting of the previously planted and late maturing rice and cotton crops. This leaves very limited time for land preparation for 'on-time' planting of wheat. 'No-tillage' technologies that reduce the turn-round time for wheat cultivation after rice and cotton have been developed, but their uptake has not been as expected.-This paper attempts to determine the farm and farmer characteristics and other socio-economic factors that influence the adoption of 'no-tillage' technologies'. Logit models were developed for the analysis undertaken. In the 'cotton-wheat' system personal characteristics like education, tenancy status, attitude towards risk implied in the use of new technologies and contact with extension agents are the main factors that affect adoption. As regards the 'rice-wheat' system, resource endowments such as farm size, access to a 'no-tillage' drill, clayey soils and the area sown to the rice-wheat sequence along with tenancy and contact with extension agents were dominant in explaining adoption. (C) 2002 Elsevier Science Ltd. All rights reserved.

Economic impact of Turnip mosaic virus, Cauliflower mosaic virus and Beet mosaic virus in three Kenyan vegetables

Screenhouse experiments conducted in Kenya showed that inoculation of cabbage seedlings with Turnip mosaic virus (TuMV), either alone, or in combination with Cauliflower mosaic virus (CaMV), reduced the number and weight of marketable harvested heads. When viruses were inoculated simultaneously, 25% of cabbage heads were non-marketable, representing 20-fold loss compared with control. By contrast, inoculation with CaMV alone had insignificant effects on cabbage yield. This suggests that TuMV is the more detrimental of these pathogens, and its management should be a priority. Early exposure to TuMV produced cabbages that were 50% lighter than non-infected plants, but later infection was less damaging suggesting that controlling virus infection at the seedling stage is more important. TuMV was far less damaging to kale than it was to cabbage; although high proportions of TuMV-inoculated kale plants showed symptoms (> 90%), the marketability and quality of leaves were not significantly reduced, and no clear relationship existed between timing of infection and subsequent crop losses. Early inoculation of Swiss chard with Beet mosaic virus (BtMV) significantly impaired leaf quality (similar to 50% reduction in marketable leaf production), but the impact of disease was greatest in plants that had been inoculated at maturity, where average leaf losses were two and a half times those recorded in virus-free plants. Disease-management of BtMV in Swiss chard is important, therefore, not only at the seedling stage, but particularly when plants are transplanted from nursery to field.

Status and distribution of the Grey-headed Fish-Eagle (Ichthyophaga ichthyaetus) in the Prek Toal Core Area of Tonle Sap Lake, Cambodia

The regional population of the Grey-headed Fish-Eagle (Ichthyophaga ichthyaetus) in Southeast Asia is thought to be in recent decline and its conservation status Linder threat. We undertook a systematic survey in a flooded swamp forest at the Tonle Sap Lake in Cambodia and recorded 32 pairs of eagles in an area of approximately 80 km(2). Three species of water snakes were identified as eagle prey items, previously unrecorded for this species. We suggest that this eagle population has significant regional importance and discuss potential anthropogenic threats to population stability, such as water snake harvesting and construction Of upstream hydropower dams.

Factors affecting biomass production and nutritive value of Calliandra calothyrsus leaf as fodder for ruminants

Calliandra calothyrsus is a tree legume native to Mexico and Central America. The species has attracted considerable attention for its capacity to produce both fuelwood and foliage for either green manure or fodder. Its high content of proanthocyanidins (condensed tannins) and associated low digestibility has, however, limited its use as a feed for ruminants, and there is also a widespread perception that wilting the leaves further reduces their nutritive value. Nevertheless, there has been increasing uptake of calliandra as fodder in certain regions, notably the Central Highlands of Kenya. The present study, conducted in Embu, Kenya, investigated effects of provenance, wilting, cutting frequency and seasonal variation both in the laboratory (in vitro digestibility, crude protein, neutral detergent fibre, extractable and bound proanthocyanidins) and in on-station animal production trials with growing lambs and lactating goats. The local Kenyan landrace of calliandra (Embu) and a closely-related Guatemalan provenance (Patulul) were found to be significantly different, and superior, to a provenance from Nicaragua (San Ramon) in most of the laboratory traits measured, as well as in animal production and feed efficiency. Cutting frequency had no important effect on quality; and although all quality traits displayed seasonal variation there was little discernible pattern to this variation. Wilting had a much less negative effect than expected, and for lambs fed calliandra as a supplement to a low quality basal feed (maize stover), wilting was actually found to give higher live-weight gain and feed efficiency. Conversely, with a high quality basal diet (Napier grass) wilting enhanced intake but not live-weight gain, so feed efficiency was greater for fresh material. The difference between fresh and wilted leaves was not great enough to justify the current widespread recommendation that calliandra should always be fed fresh.

Floral volatiles controlling ant behaviour

P>1. Ants show complex interactions with plants, both facultative and mutualistic, ranging from grazers through seed predators and dispersers to herders of some herbivores and guards against others. But ants are rarely pollinators, and their visits to flowers may be detrimental to plant fitness. 2. Plants therefore have various strategies to control ant distributions, and restrict them to foliage rather than flowers. These 'filters' may involve physical barriers on or around flowers, or 'decoys and bribes' sited on the foliage (usually extrafloral nectaries - EFNs). Alternatively, volatile organic compounds (VOCs) are used as signals to control ant behaviour, attracting ants to leaves and/or deterring them from functional flowers. Some of the past evidence that flowers repel ants by VOCs has been equivocal and we describe the shortcomings of some experimental approaches, which involve behavioural tests in artificial conditions. 3. We review our previous study of myrmecophytic acacias, which used in situ experiments to show that volatiles derived from pollen can specifically and transiently deter ants during dehiscence, the effects being stronger in ant-guarded species and more effective on resident ants, both in African and Neotropical species. In these plants, repellence involves at least some volatiles that are known components of ant alarm pheromones, but are not repellent to beneficial bee visitors. 4. We also present new evidence of ant repellence by VOCs in temperate flowers, which is usually pollen-based and active on common European ants. We use these data to indicate that across a wide range of plants there is an apparent trade-off in ant-controlling filter strategies between the use of defensive floral volatiles and the alternatives of decoying EFNs or physical barriers.

Grazing management of calcareous grasslands and its implications for the conservation of beetle communities

Calcareous grasslands are an important habitat for floral and faunal communities in the UK and Europe. Declines due to changes in management, scrub invasion and agricultural improvement have left much of the remnants of this habitat in a degraded and fragmented state. Grazing, by cattle or sheep, is one of the main management practices used to maintain and improve the floral and faunal quality of calcareous grassland. The long-term impacts of different grazing regimes, however, are poorly understood, particularly in terms of the invertebrate communities. This study contrasted the impacts of recently introduced and long-term sheep or cattle grazing on beetle communities present on one of the largest areas of calcareous grassland in Europe, the Salisbury Plain military training Area, UK. No effects of grazing management on beetle abundance, species. richness or evenness were found, but plant diversity and overall percentage cover of grasses did influence beetle diversity. Proportions of the total number of individuals and overall species richness within beetle guilds (predatory, phytophagous, flower/seed feeders, root feeders and foliage feeders) were strongly influenced by both the duration and type of grazing animal. At the species level, beetle community structure showed significant differences between ungrazed, long-term cattle and long-term sheep grazing treatments. Changes in plant community structure were found to influence beetle community structure. The significance of these results is discussed in terms of the long-term impacts of grazing on beetle community structure, and the benefits of different grazing regimes for the conservation management of calcareous grasslands. (c) 2005 Elsevier Ltd. All rights reserved.

Shoot dieback in clipped young Golden Leyland (Cupressocyparis leylandii) trees - a physiological mechanism?

Shoot dieback is a problem in frequently trimmed Leyland hedges and is increasingly affecting gardeners’ choice of hedge trees, having a negative effect on a conifer nursery industry. Some damage can be attributed to the feeding by aphids, but it is unclear if there are also underlying physiological causes. In this study, we tested the hypothesis that shoot-clipping of conifer trees during adverse growing conditions (i.e. high air temperature and low soil moisture) could be leading to shoot ‘dieback’. Three-year-old Golden Leyland Cypress (x Cupressocyparis leylandii ‘Excalibur Gold’) plants were subjected to either a well-watered or droughted irrigation regime and placed in either a ‘hot’ (average day temperature = 40°C) or a ‘cool’ (average day temperature = 27°C) glasshouse compartment. Half of the plants from each glasshouse were clipped on Day 14 and again on Day 50. Measurements of soil moisture content (SMC), net CO2 assimilation rate (A), stomatal conductance (gs), branchlet xylem water potential (XWP), plant height and foliage colour were made. Within the clipped and unclipped treatments of both glasshouse compartments, plants from the droughted regime had significantly lower values for A, gs and XWP than those from the well-watered regime. However, there was no difference in these parameters between the hot and cool glasshouse compartments. The trends seen for A, gs and XWP of all treatments generally mirrored changes in SMC indicating a direct effect of water supply on these parameters. By the end of the experiment the overall foliage colour of plants from the hot glasshouse was darker than that of plants from the cool glasshouse and the overall foliage colour was also darker following shoot clipping. In general, shoot clipping led to increases in A, gs XWP and SMC. This may be due to the reduction in total leaf area leading to a greater supply of water for the remaining leaves. No shoot ‘dieback’ was observed in any treatment in response to drought stress or shoot-clipping.

The use of image analysis to estimate population growth rate in Daphnia magna

1. Population growth rate (PGR) is central to the theory of population ecology and is crucial for projecting population trends in conservation biology, pest management and wildlife harvesting. Furthermore, PGR is increasingly used to assess the effects of stressors. Image analysis that can automatically count and measure photographed individuals offers a potential methodology for estimating PGR. 2. This study evaluated two ways in which the PGR of Daphnia magna, exposed to different stressors, can be estimated using an image analysis system. The first method estimated PGR as the ratio of counts of individuals obtained at two different times, while the second method estimated PGR as the ratio of population sizes at two different times, where size is measured by the sum of the individuals' surface areas, i.e. total population surface area. This method is attractive if surface area is correlated with reproductive value (RV), as it is for D. magna, because of the theoretical result that PGR is the rate at which the population RV increases. 3. The image analysis system proved reliable and reproducible in counting populations of up to 440 individuals in 5 L of water. Image counts correlated well with manual counts but with a systematic underestimate of about 30%. This does not affect accuracy when estimating PGR as the ratio of two counts. Area estimates of PGR correlated well with count estimates, but were systematically higher, possibly reflecting their greater accuracy in the study situation. 4. Analysis of relevant scenarios suggested the correlation between RV and body size will generally be good for organisms in which fecundity correlates with body size. In these circumstances, area estimation of PGR is theoretically better than count estimation. 5. Synthesis and applications. There are both theoretical and practical advantages to area estimation of population growth rate when individuals' reproductive values are consistently well correlated with their surface areas. Because stressors may affect both the number and quality of individuals, area estimation of population growth rate should improve the accuracy of predicting stress impacts at the population level.

Growth, yield and development of strawberry cv. 'Elsanta' under novel photoselective film clad greenhouses

Film greenhouse claddings are typically used to protect horticultural crops from low temperature and high rainfall conditions. However, a range of novel plastic films have been developed which filter specific radiation wavelengths with the aim of providing the grower with greater control of crop growth and development. A replicated experiment was conducted in 8 small experimental greenhouses, covered with different photoselective films characterised by a range of red/far-red ratios and PAR transmissions to study their effects on the growth, yield and quality of the strawberry cv. 'Elsanta'. Marketable yield per plant was 51% greater under the film with the highest light transmission (control) compared with the lower light transmission films. Similarly, unmarketable fruit number and average non-marketable individual fruit weight per plant was lowest under films with higher light transmissions and higher under lower light transmission films. Cropping duration was longer under films with high R/FR. Also plants under high R/FR were more compact (due to shorter petiole lengths) compared to plants grown under low R/FR. The results are discussed in relation to the increased use of photoselective films in protected horticulture and the need for higher quality fruit and vegetables.

Influence of harvest date and crop yield on the fatty acid composition of virgin olive oils from cv. Picual

In this study was analyzed the effect of crop year and harvesting time on the fatty acid composition of cv. Picual virgin olive oil. The study was carried out during the fruit ripening period for three crop seasons. The mean fatty acid composition of Picual oils was determined. The oils contained palmitic acid (11.9%), oleic acid (79.3%), and linoleic acid (2.95%). The content of palmitic acid and saturated fatty acids decreased during fruit ripening while oleic and linoleic acids increased. The amount of stearic and linolenic acids decreased. The amount of saturated acids, palmitic and stearic, and the polyunsaturated acids linoleic and linolenic was dependent on the time of harvest, whereas the amount of oleic acid varied with the crop year. The differences observed between crop years for both palmitic and linoleic acid may be explained by the differences in the temperature during oil biosynthesis and by the amount of summer rainfall for oleic acid content. A significant relationship was observed between the MUFA/PUFA ratio and the oxidative stability measured by the Rancimat method.

Seasonal changes in olive fruit characteristics and oil accumulation during ripening process

Olive fruits of three of the most important Spanish and Italian cultivars, 'Picual', `Hojiblanca' and 'Frantoio', were harvested at bi-weekly periods during three crop seasons to study their development and ripening process. Fresh and dry weights and ripening index were determined for fruit, while dry matter, oil and moisture contents were determined in both fruit and pulp (flesh). Fruit growth rate and oil accumulation were calculated. Each olive cultivar showed a different ripening pattern, 'Hojiblanca' being the last one to maturate. Fruit weight increased, decreasing its growth rate from the middle of November. Dry matter and moisture contents decreased during ripening in pulp and fruit, 'Hojiblanca' showing the highest values for both. Oil content, when expressed on a fresh weight basis, increased in all cultivars, although for the last time period showed variations due to climatic conditions. During ripening, oil content on a dry weight basis increased in fruit, but oil biosynthesis in flesh ceased from November. Olive fruits presented lower oil and higher dry matter contents in the year of lowest rainfall. Therefore fruit harvesting should be carried out from the middle of November in order to obtain the highest oil yield and avoid natural fruit drop. (C) 2004 Society of Chemical Industry.

ontoReadiness: A meta-ontology for readiness certification and career portability

This paper argues the need for the information communication technology (ICT), labor exchange (job boards), and Human Capital ontology engineers (ontoEngineers) to jointly design and socialize an upper level meta-ontology for people readiness and career portability. These enticing ontology research topics have yielded "independent" results, but have yet to meet the more broader or "universal" requirement that emerging frameworks demand. This paper will focus on the need to universally develop an upper level ontology and provide the reader concepts and models that can be transformed into marketable solutions.

Bio-energy retains its mitigation potential under elevated CO2

Background If biofuels are to be a viable substitute for fossil fuels, it is essential that they retain their potential to mitigate climate change under future atmospheric conditions. Elevated atmospheric CO2 concentration [CO2] stimulates plant biomass production; however, the beneficial effects of increased production may be offset by higher energy costs in crop management. Methodology/Main findings We maintained full size poplar short rotation coppice (SRC) systems under both current ambient and future elevated [CO2] (550 ppm) and estimated their net energy and greenhouse gas balance. We show that a poplar SRC system is energy efficient and produces more energy than required for coppice management. Even more, elevated [CO2] will increase the net energy production and greenhouse gas balance of a SRC system with 18%. Managing the trees in shorter rotation cycles (i.e. 2 year cycles instead of 3 year cycles) will further enhance the benefits from elevated [CO2] on both the net energy and greenhouse gas balance. Conclusions/significance Adapting coppice management to the future atmospheric [CO2] is necessary to fully benefit from the climate mitigation potential of bio-energy systems. Further, a future increase in potential biomass production due to elevated [CO2] outweighs the increased production costs resulting in a northward extension of the area where SRC is greenhouse gas neutral. Currently, the main part of the European terrestrial carbon sink is found in forest biomass and attributed to harvesting less than the annual growth in wood. Because SRC is intensively managed, with a higher turnover in wood production than conventional forest, northward expansion of SRC is likely to erode the European terrestrial carbon sink.