Latitudinal gradients in climatic‐niche evolution accelerate trait evolution at high latitudes

Despite the importance of divergent selection to the speed of evolution, it remains poorly understood if divergent selection is more prevalent in the tropics (where species richness is highest), or at high latitudes (where paleoclimate change has been most intense). We tested whether the rate of climatic‐niche evolution – one proxy for divergent selection – varies with latitude for 111 pairs of bird species. Using Brownian motion and Ornsetin–Ulhenbeck models, we show that evolutionary rates along two important axes of the climatic‐niche – temperature and seasonality – have been faster at higher latitudes. We then tested whether divergence of the climatic‐niche was associated with evolution in traits important in ecological differentiation (body mass) and reproductive isolation (song), and found that climatic divergence is associated with faster rates in both measures. These results highlight the importance of climate‐mediated divergent selection pressures in driving evolutionary divergence and reproductive isolation at high latitudes.     

Molecular approaches to diversity of populations of apicomplexan parasites

Apicomplexan parasites include many parasites of importance either for livestock or as causative agents of human diseases. The importance of these parasites has been recognised by the European Commission and resulted in support of the COST (Cooperation in Science and Technology) Action 857 ‘Apicomplexan Biology in the Post-Genomic Era’. In this review we discuss the current understanding in ‘Biodiversity and Population Genetics’ of the major apicomplexan parasites, namely the Eimeria spp., Cryptosporidium spp., Toxoplasma gondii, Neosporacaninum, Theileria spp. and Plasmodium spp. During the past decade molecular tools for characterizing and monitoring parasite populations have been firmly established as an integral part of field studies and intervention trials. Analyses have been conducted for most apicomplexan pathogens to describe the extent of genetic diversity, infection dynamics or population structure. The underlying key question for all parasites is to understand how genetic diversity influences epidemiology and pathogenicity and its implication in therapeutic and vaccination strategies as well as disease control. Similarities in the basic biology and disease or transmission patterns among this order of parasites promote multifaceted discussions and comparison of epidemiological approaches and methodological tools. This fosters mutual learning and has the potential for cross-fertilisation of ideas and technical approaches.     

Evaluation of the Cepheid GeneXpert system for detecting Bacillus anthracis

AIMS: The Cepheid GeneXpert is a four-site, automated sample preparation and real-time PCR detection system. In this study, the capability of the GeneXpert to isolate and detect nucleic acid from Bacillus anthracis Ames spores was assessed. METHODS AND RESULTS: A four-plex, dried-down bead cartridge containing PCR reagents specific for the pXO1 and pXO2 plasmids as well as sample processing and inhibition controls was evaluated. For B. anthracis Ames spores harbouring pXO1 and pXO2, samples containing 68 CFU per ml (148 spores per ml) were positive in all four replicates. A limited cross-reactivity panel, which included closely related Bacillus species, was also tested to determine the specificity of the pXO1 and pXO2 assays. No cross-reactivity occurred. Further, B. anthracis Sterne spore samples were analysed to compare results when processed using the GeneXpert to those run directly on the Cepheid SmartCycler without sample processing. The GeneXpert detection capability was three logs lower than the SmartCycler indicating the benefit of incorporating a nucleic acid extraction procedure. CONCLUSIONS: This study demonstrates that the GeneXpert is a rapid and reliable system for simultaneously detecting the B. anthracis virulence plasmids pXO1 and pXO2. SIGNIFICANCE AND IMPACT OF THE STUDY: The GeneXpert is the only platform currently available that is capable of both nucleic acid purification and real-time PCR detection enclosed within a single system. Further, all sample manipulations are automated, thus reducing errors associated with manual processing.     

Quantitative identification of fecal water pollution sources by TaqMan real-time PCR assays using Bacteroidales 16S rRNA genetic markers

PCR-based analysis of Bacteroidales 16S rRNA genes has emerged as a promising tool to identify sources of fecal water pollution. In this study, three TaqMan real-time PCR assays (BacGeneral, BacHuman, and BacBovine) were developed and evaluated for their ability to quantitatively detect general (total), human-specific, and bovine-specific Bacteroidales 16S rRNA genetic markers. The detection sensitivity was determined to be 6.5 copies of 16S rRNA gene for the BacGeneral and BacHuman assays and 10 copies for the BacBovine assay. The assays were capable of detecting approximately one to two cells per PCR. When tested with 70 fecal samples from various sources (human, cattle, pig, deer, dog, cat, goose, gull, horse, and raccoon), the three assays positively identified the target markers in all samples without any false-negative results. The BacHuman and BacBovine assays exhibited false-positive reactions with non-target samples in a few cases. However, the level of the false-positive reactions was about 50 times smaller than that of the true-positive ones, and therefore, these cross-reactions were unlikely to cause misidentifications of the fecal pollution sources. Microbial source-tracking capability was tested at two freshwater streams of which water quality was influenced by human and cattle feces, respectively. The assays accurately detected the presence of the corresponding host-specific markers upon fecal pollution and the persistence of the markers in downstream areas. The assays are expected to reliably determine human and bovine fecal pollution sources in environmental water samples.     

Statistical genetic issues for genome-wide association studies

Genotyping technology now allows the rapid and affordable generation of million-SNP profiles for humans, leading to considerable activity in association mapping. Similar activity is anticipated for many plant species, including Brassica. These plant association mapping activities will require the same care in quality control and quality assurance as for humans. The subsequent analyses may draw upon the same body of theory that is described here in the language of quantitative genetics.     

Mechanomyographic and electromyographic responses of the vastus medialis muscle during isometric and concentric muscle actions

The purpose of this study was to examine the patterns for the mechanomyographic (MMG) and electromyographic (EMG) amplitude and mean power frequency (MPF) vs. torque relationships during submaximal to maximal isometric and isokinetic muscle actions. Seven men (mean +/- SD age, 22.4 +/- 1.3 years) volunteered to perform isometric and concentric isokinetic leg extension muscle actions at 20, 40, 60, 80, and 100% of maximal voluntary contraction (MVC) and peak torque (PT) on a Cybex II dynamometer. A piezoelectric MMG recording sensor was placed between bipolar surface EMG electrodes on the vastus medialis. Polynomial regression and separate 1-way repeated-measures analysis of variance were used to analyze the EMG amplitude, MMG amplitude, EMG MPF, and MMG MPF data for the isometric and isokinetic muscle actions. For the isometric muscle actions, EMG amplitude (R(2) = 0.999) and MMG MPF (R(2) = 0.946) increased to MVC, mean MMG amplitude increased to 60% MVC and then plateaued, and mean EMG MPF did not change (p > 0.05) across torque levels. For the isokinetic muscle actions, EMG amplitude (R(2) = 0.988) and MMG amplitude (R(2) = 0.933) increased to PT, but there were no significant mean changes with torque for EMG MPF or MMG MPF. The different torque-related responses for EMG and MMG amplitude and MPF may reflect differences in the motor control strategies that modulate torque production for isometric vs. dynamic muscle actions. These results support the findings of others and suggest that isometric torque production was modulated by a combination of recruitment and firing rate, whereas dynamic torque production was modulated primarily through recruitment.     

ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis

Nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) are key signalling molecules produced in response to various stimuli and involved in a diverse range of plant signal transduction processes. Nitric oxide and H(2)O(2) have been identified as essential components of the complex signalling network inducing stomatal closure in response to the phytohormone abscisic acid (ABA). A close inter-relationship exists between ABA and the spatial and temporal production and action of both NO and H(2)O(2) in guard cells. This study shows that, in Arabidopsis thaliana guard cells, ABA-mediated NO generation is in fact dependent on ABA-induced H(2)O(2) production. Stomatal closure induced by H(2)O(2) is inhibited by the removal of NO with NO scavenger, and both ABA and H(2)O(2) stimulate guard cell NO synthesis. Conversely, NO-induced stomatal closure does not require H(2)O(2) synthesis nor does NO treatment induce H(2)O(2) production in guard cells. Tungstate inhibition of the NO-generating enzyme nitrate reductase (NR) attenuates NO production in response to nitrite in vitro and in response to H(2)O(2) and ABA in vivo. Genetic data demonstrate that NR is the major source of NO in guard cells in response to ABA-mediated H(2)O(2) synthesis. In the NR double mutant nia1, nia2 both ABA and H(2)O(2) fail to induce NO production or stomatal closure, but in the nitric oxide synthase deficient Atnos1 mutant, responses to H(2)O(2) are not impaired. Importantly, we show that in the NADPH oxidase deficient double mutant atrbohD/F, NO synthesis and stomatal closure to ABA are severely reduced, indicating that endogenous H(2)O(2) production induced by ABA is required for NO synthesis. In summary, our physiological and genetic data demonstrate a strong inter-relationship between ABA, endogenous H(2)O(2) and NO-induced stomatal closure.     

Gender comparisons of mechanomyographic amplitude and mean power frequency versus isometric torque relationships

This study compared the patterns of mechanomyographic (MMG) amplitude and mean power frequency vs. torque relationships in men and women during isometric muscle actions of the biceps brachii. Seven men (mean age 23.9 +/- 3.5 yrs) and 8 women (mean 21.0 +/- 1.3 yrs) performed submaximal to maximal isometric muscle actions of the dominant forearm flexors. Following determination of the isometric maximum voluntary contraction (MVC), they randomly performed submaximal step muscle actions in 10% increments from 10% to 90% MVC. Polynomial regression analyses indicated that the MMG amplitude vs. isometric torque relationship for the men was best fit with a cubic model (R(2) = 0.983),,where MMG amplitude increased slightly from 10% to 20% MVC, increased rapidly from 20% to 80% MVC, and plateaued from 80% to 100% MVC. For the women, MMG amplitude increased linearly (r(2) = 0.949) from 10% to 100% MVC. Linear models also provided the best fit for the MMG mean power frequency vs. isometric torque relationship in both the men (r(2) = 0.813) and women (r(2) = 0.578). The results demonstrated gender differences in the MMG amplitude vs. isometric torque relationship, but similar torque-related patterns for MMG mean power frequency. These findings suggested that the plateau in MMG amplitude at high levels of isometric torque production for the biceps brachii in the men, but not the women, may have been due to greater isometric torque, muscle stiffness, and/or intramuscular fluid pressure in the men, rather than to differences in motor unit activation strategies for modulating isometric torque production.