Excess polymorphisms in genes for membrane proteins in Plasmodium falciparum

The detection of single-nucleotide polymorphisms in pathogenic microorganisms has normally been carried out by trial and error. Here we show that DNA hybridization with high-density oligonucleotide arrays provides rapid and convenient detection of single-nucleotide polymorphisms in Plasmodium falciparum, despite its exceptionally high adenine-thymine (AT) content (82%). A disproportionate number of polymorphisms are found in genes encoding proteins associated with the cell membrane. These genes are targets for only 22% of the oligonucleotide probes but account for 69% of the polymorphisms. Genetic variation is also enriched in subtelomeric regions, which account for 22% of the chromosome but 76% of the polymorphisms.     

Discovery of gene function by expression profiling of the malaria parasite life cycle

The completion of the genome sequence for Plasmodium falciparum, the species responsible for most malaria human deaths, has the potential to reveal hundreds of new drug targets and proteins involved in pathogenesis. However, only approximately 35% of the genes code for proteins with an identifiable function. The absence of routine genetic tools for studying Plasmodium parasites suggests that this number is unlikely to change quickly if conventional serial methods are used to characterize encoded proteins. Here, we use a high-density oligonucleotide array to generate expression profiles of human and mosquito stages of the malaria parasite's life cycle. Genes with highly correlated levels and temporal patterns of expression were often involved in similar functions or cellular processes.     

Linking changes in the soil microbial community to C and N dynamics during crop residue decomposition

Crop residues are among the main inputs that allow the organic carbon (C) and nutrients to be maintained in agricultural soil. It is an important management strategy that can improve soil fertility and enhance agricultural productivity. This work aims to evaluate the extent of the changes that may occur in the soil heterotrophic microbial communities involved in organic matter decomposition and C and nitrogen (N) mineralization after the addition of crop residues. Soil microcosm experiments were performed at 28°C for 90 days with the addition of three crop residues with contrasting biochemical qualities: pea (Pisum sativum L.), rapeseed (Brassica napus L.), and wheat (Triticum aestivum L.). Enzyme activities, C and N mineralization, and bacterial and fungal biomasses were monitored, along with the bacterial and fungal community composition, by the high-throughput sequencing of 16S rRNA and ITS genes. The addition of crop residues caused decreases in β-glucosidase and arylamidase activities and simultaneous enhancement of the C mineralization and net N immobilization, which were linked to changes in the soil microbial communities. The addition of crop residues decreased the bacterial and fungal biomasses 90 days after treatment and there were shifts in bacterial and fungal diversity at the phyla, order, and genera levels. Some specific orders and genera were dependent on crop residue type. For example, Chloroflexales, Inquilinus, Rubricoccus, Clitocybe, and Verticillium were identified in soils with pea residues; whereas Thermoanaerobacterales, Thermacetogenum, and Hypoxylon were enriched in soils with rapeseed residues, and Halanaerobiales, Rubrobacter, and Volutella were only present in soils with wheat residues. The findings of this study suggest that soil C and N dynamics in the presence of the crop residues were driven by the selection of specific bacterial and fungal decomposers linked to the biochemical qualities of the crop residues. If crop residue decomposition processes showed specific bacterial and fungal operational taxonomic unit (OTU) signatures, this study also suggests a strong functional redundancy that exists among soil microbial communities.     

Development of a xylazine constant rate infusion with or without butorphanol for standing sedation of horses

ObjectiveTo elaborate constant rate infusion (CRI) protocols for xylazine (X) and xylazine/butorphanol (XB) which will result in constant sedation and steady xylazine plasma concentrations.Study designBlinded randomized experimental study.AnimalsTen adult research horses.MethodsPart I: After normal height of head above ground (HHAG = 100%) was determined, a loading dose of xylazine (1 mg kg^?1) with butorphanol (XB: 18 μg kg^?1) or saline (X: equal volume) was given slowly intravenously (IV). Immediately afterwards, a CRI of butorphanol (XB: 25 μg kg^?1 hour^?1) or saline (X) was administered for 2 hours. The HHAG was used as a marker of depth of sedation. Sedation was maintained for 2 hours by additional boluses of xylazine (0.3 mg kg^?1) whenever HHAG >50%. The dose of xylazine (mg kg^?1 hour^?1) required to maintain sedation was calculated for both groups. Part II: After the initial loading dose, the calculated xylazine infusion rates were administered in parallel to butorphanol (XB) or saline (X) and sedation evaluated. Xylazine plasma concentrations were measured by HPLC-MS-MS at time points 0, 5, 30, 45, 60, 90, and 120 minutes. Data were analyzed using paired t-test, Wilcoxon signed rank test and a 2-way anova for repeated measures (p < 0.05).ResultsThere was no significant difference in xylazine requirements (X: 0.69, XB: 0.65 mg kg^?1 hour^?1) between groups. With treatment X, a CRI leading to prolonged sedation was developed. With XB, five horses (part I: two, part II: three) fell down and during part II four horses appeared insufficiently sedated. Xylazine plasma concentrations were constant after 45 minutes in both groups.ConclusionXylazine bolus, followed by CRI, provided constant sedation. Additional butorphanol was ineffective in reducing xylazine requirements and increased ataxia and apparent early recovery from sedation in unstimulated horses.Clinical relevanceData were obtained on unstimulated healthy horses and extrapolation to clinical conditions requires caution.     

Identifying and mapping biodiversity processes for conservation planning in islands: A case study in Réunion Island (Western Indian Ocean)

Over the last century, island biodiversity has become one of the most threatened in the world. Although many island conservation plans address biodiversity requirements at the species level, few plans address the spatial requirements of the biodiversity processes that underpin the persistence of these species. Using systematic conservation planning principles, we map the spatial components of biodiversity processes (SCBPs) and use these to design broad-scale conservation corridors for Réunion Island. Our method is based upon a literature review, expert knowledge, spatially explicit base data, conservation planning software, and spatial modelling. We combine a target-driven algorithm with least-cost path analyses to delineate optimal corridors for capturing key biodiversity processes while simultaneously considering biodiversity pattern targets, conservation opportunities, and future threats. We identify five SCBPs: the oceanic-terrestrial interface; riverine corridors; macrohabitat interfaces; the boundaries of isolated topographic units; and lowland-upland gradients. A large proportion of the SCBPs (81.3%) is currently untransformed, whereas 3% is irreversibly transformed by urbanisation and 15.7% is transformed but restorable. However, SCBPs are almost fully disrupted by urbanisation in the lowlands, thereby compromising functional corridors along full altitudinal gradients. This study is a contribution toward the reconciliation of conservation versus development objectives on Réunion Island but we believe that the delineation method is sufficiently general to be applied to other islands. Our results highlight the need for integrating marine, coastal and terrestrial conservation planning as a matter of urgency, given the rapid transformation of coastal areas on islands.