Rhipicephalus microplus serine protease inhibitor family: annotation, expression and functional characterisation assessment

Background: Rhipicephalus (Boophilus) microplus evades the host's haemostatic system through a complex protein array secreted into tick saliva. Serine protease inhibitors (serpins) conform an important component of saliva which are represented by a large protease inhibitor family in Ixodidae. These secreted and non-secreted inhibitors modulate diverse and essential proteases involved in different physiological processes. Methods: The identification of R. microplus serpin sequences was performed through a web-based bioinformatics environment called Yabi. The database search was conducted on BmiGi V1, BmiGi V2.1, five SSH libraries, Australian tick transcriptome libraries and RmiTR V1 using bioinformatics methods. Semi quantitative PCR was carried out using different adult tissues and tick development stages. The cDNA of four identified R. microplus serpins were cloned and expressed in Pichia pastoris in order to determine biological targets of these serpins utilising protease inhibition assays. Results: A total of four out of twenty-two serpins identified in our analysis are new R. microplus serpins which were named as RmS-19 to RmS-22. The analyses of DNA and predicted amino acid sequences showed high conservation of the R. microplus serpin sequences. The expression data suggested ubiquitous expression of RmS except for RmS-6 and RmS-14 that were expressed only in nymphs and adult female ovaries, respectively. RmS-19, and -20 were expressed in all tissues samples analysed showing their important role in both parasitic and non-parasitic stages of R. microplus development. RmS-21 was not detected in ovaries and RmS-22 was not identified in ovary and nymph samples but were expressed in the rest of the samples analysed. A total of four expressed recombinant serpins showed protease specific inhibition for Chymotrypsin (RmS-1 and RmS-6), Chymotrypsin / Elastase (RmS-3) and Thrombin (RmS-15). Conclusion: This study constitutes an important contribution and improvement to the knowledge about the physiologic role of R. microplus serpins during the host-tick interaction.

Assessment of functional forms of crop yield loss models of invasive plant species applied in decision support tools and bioeconomic modelling

AbstractObjectives Decision support tools (DSTs) for invasive species management have had limited success in producing convincing results and meeting users' expectations. The problems could be linked to the functional form of model which represents the dynamic relationship between the invasive species and crop yield loss in the DSTs. The objectives of this study were: a) to compile and review the models tested on field experiments and applied to DSTs; and b) to do an empirical evaluation of some popular models and alternatives. Design and methods This study surveyed the literature and documented strengths and weaknesses of the functional forms of yield loss models. Some widely used models (linear, relative yield and hyperbolic models) and two potentially useful models (the double-scaled and density-scaled models) were evaluated for a wide range of weed densities, maximum potential yield loss and maximum yield loss per weed. Results Popular functional forms include hyperbolic, sigmoid, linear, quadratic and inverse models. Many basic models were modified to account for the effect of important factors (weather, tillage and growth stage of crop at weed emergence) influencing weed–crop interaction and to improve prediction accuracy. This limited their applicability for use in DSTs as they became less generalized in nature and often were applicable to a much narrower range of conditions than would be encountered in the use of DSTs. These factors' effects could be better accounted by using other techniques. Among the model empirically assessed, the linear model is a very simple model which appears to work well at sparse weed densities, but it produces unrealistic behaviour at high densities. The relative-yield model exhibits expected behaviour at high densities and high levels of maximum yield loss per weed but probably underestimates yield loss at low to intermediate densities. The hyperbolic model demonstrated reasonable behaviour at lower weed densities, but produced biologically unreasonable behaviour at low rates of loss per weed and high yield loss at the maximum weed density. The density-scaled model is not sensitive to the yield loss at maximum weed density in terms of the number of weeds that will produce a certain proportion of that maximum yield loss. The double-scaled model appeared to produce more robust estimates of the impact of weeds under a wide range of conditions. Conclusions Previously tested functional forms exhibit problems for use in DSTs for crop yield loss modelling. Of the models evaluated, the double-scaled model exhibits desirable qualitative behaviour under most circumstances.

Resource use and greenhouse gas emissions from grain-finishing beef cattle in seven Australian feedlots: a life cycle assessment

Grain finishing of cattle has become increasingly common in Australia over the past 30 years. However, interest in the associated environmental impacts and resource use is increasing and requires detailed analysis. In this study we conducted a life cycle assessment (LCA) to investigate impacts of the grain-finishing stage for cattle in seven feedlots in eastern Australia, with a particular focus on the feedlot stage, including the impacts from producing the ration, feedlot operations, transport, and livestock emissions while cattle are in the feedlot (gate-to-gate). The functional unit was 1 kg of liveweight gain (LWG) for the feedlot stage and results are included for the full supply chain (cradle-to-gate), reported per kilogram of liveweight (LW) at the point of slaughter. Three classes of cattle produced for different markets were studied: short-fed domestic market (55–80 days on feed), mid-fed export (108–164 days on feed) and long-fed export (>300 days on feed). In the feedlot stage, mean fresh water consumption was found to vary from 171.9 to 672.6 L/kg LWG and mean stress-weighted water use ranged from 100.9 to 193.2 water stress index eq. L/kg LWG. Irrigation contributed 57–91% of total fresh water consumption with differences mainly related to the availability of irrigation water near the feedlot and the use of irrigated feed inputs in rations. Mean fossil energy demand ranged from 16.5 to 34.2 MJ lower heating values/kg LWG and arable land occupation from 18.7 to 40.5 m2/kg LWG in the feedlot stage. Mean greenhouse gas (GHG) emissions in the feedlot stage ranged from 4.6 to 9.5 kg CO2-e/kg LWG (excluding land use and direct land-use change emissions). Emissions were dominated by enteric methane and contributions from the production, transport and milling of feed inputs. Linear regression analysis showed that the feed conversion ratio was able to explain >86% of the variation in GHG intensity and energy demand. The feedlot stage contributed between 26% and 44% of total slaughter weight for the classes of cattle fed, whereas the contribution of this phase to resource use varied from 4% to 96% showing impacts from the finishing phase varied considerably, compared with the breeding and backgrounding. GHG emissions and total land occupation per kilogram of LWG during the grain finishing phase were lower than emissions from breeding and backgrounding, resulting in lower life-time emissions for grain-finished cattle compared with grass finishing.