Control of the phytopathogen Botrytis cinerea using adipic acid monoethyl ester

The in vitro and in vivo antifungal activity of adipic acid monoethyl ester (AAME) on the necrotrophic pathogen Botrytis cinerea has been studied. This chemical effectively controlled this important phytopathogen, inhibited spore germination and mycelium development at non-phytotoxic concentrations. The effectiveness of AAME treatment is concentration-dependent and influenced by pH. Spore germination in the presence of AAME is stopped at a very early stage, preventing germ tube development. In addition, cytological changes such as retraction of the conidial cytoplasm in the fungus are observed. AAME was also found to act on membrane integrity, affecting permeability without exhibiting lytic activity, as described previously for other antifungal compounds. Polyamine content in the mycelium of B. cinerea was also affected in response to AAME treatment, resulting in putrescine reduction and spermine accumulation similar to a number of antifungal agents. Microscopic observation of treated conidia after inoculation on tomato leaves suggested that inhibited spores are not able to attach to and penetrate the leaf. Finally, AAME completely suppressed the grey mould disease of tomato fruits under controlled inoculation conditions, providing evidence for its efficacy in a biological context and for the potential use of this chemical as an alternative fungicide treatment.     

Localization of the Bacillus subtilis murB gene within the dcw cluster is important for growth and sporulation

The Bacillus subtilis murB gene, encoding UDP-N-acetylenolpyruvoylglucosamine reductase, a key enzyme in the peptidoglycan (PG) biosynthetic pathway, is embedded in the dcw (for "division and cell wall") cluster immediately upstream of divIB. Previous attempts to inactivate murB were unsuccessful, suggesting its essentiality. Here we show that the cell morphology, growth rate, and resistance to cell wall-active antibiotics of murB conditional mutants is a function of the expression level of murB. In one mutant, in which murB was insertionally inactivated in a merodiploid bearing a second xylose-inducible PxylA-murB allele, DivIB levels were reduced and a normal growth rate was achieved only if MurB levels were threefold that of the wild-type strain. However, expression of an extra copy of divIB restored normal growth at wild-type levels of MurB. In contrast, DivIB levels were normal in a second mutant containing an in-frame deletion of murB (DeltamurB) in the presence of the PxylA-murB gene. Furthermore, this strain grew normally with wild-type levels of MurB. During sporulation, the levels of MurB were highest at the time of synthesis of the spore cortex PG. Interestingly, the DeltamurB PxylA-murB mutant did not sporulate efficiently even at high concentrations of inducer. Since high levels of inducer did not interfere with sporulation of a murB(+)PxylA-murB strain, it appears that ectopic expression of murB fails to support efficient sporulation. These data suggest that coordinate expression of divIB and murB is important for growth and sporulation. The genetic context of the murB gene within the dcw cluster is unique to the Bacillus group and, taken together with our data, suggests that in these species it contributes to the optimal expression of cell division and PG biosynthetic functions during both vegetative growth and spore development.     

Infectious salmon anaemia virus (ISAV) isolated from the ISA disease outbreaks in Chile diverged from ISAV isolates from Norway around 1996 and was disseminated around 2005, based on surface glycoprotein gene sequences

Background

All viruses in the family Bunyaviridae possess a tripartite genome, consisting of a small, a medium, and a large RNA segment. Bunyaviruses therefore possess considerable evolutionary potential, attributable to both intramolecular changes and to genome segment reassortment. Hantaviruses (family Bunyaviridae, genus Hantavirus) are known to cause human hemorrhagic fever with renal syndrome or hantavirus pulmonary syndrome. The primary reservoir host of Sin Nombre virus is the deer mouse (Peromyscus maniculatus), which is widely distributed in North America. We investigated the prevalence of intramolecular changes and of genomic reassortment among Sin Nombre viruses detected in deer mice in three western states.

Methods

Portions of the Sin Nombre virus small (S) and medium (M) RNA segments were amplified by RT-PCR from kidney, lung, liver and spleen of seropositive peromyscine rodents, principally deer mice, collected in Colorado, New Mexico and Montana from 1995 to 2007. Both a 142 nucleotide (nt) amplicon of the M segment, encoding a portion of the G2 transmembrane glycoprotein, and a 751 nt amplicon of the S segment, encoding part of the nucleocapsid protein, were cloned and sequenced from 19 deer mice and from one brush mouse (P. boylii), S RNA but not M RNA from one deer mouse, and M RNA but not S RNA from another deer mouse.

Results

Two of 20 viruses were found to be reassortants. Within virus sequences from different rodents, the average rate of synonymous substitutions among all pair-wise comparisons (π_s) was 0.378 in the M segment and 0.312 in the S segment sequences. The replacement substitution rate (π_a) was 7.0 × 10^-4 in the M segment and 17.3 × 10^-4 in the S segment sequences. The low π_a relative to π_s suggests strong purifying selection and this was confirmed by a Fu and Li analysis. The absolute rate of molecular evolution of the M segment was 6.76 × 10^-3 substitutions/site/year. The absolute age of the M segment tree was estimated to be 37 years. In the S segment the rate of molecular evolution was 1.93 × 10^-3 substitutions/site/year and the absolute age of the tree was 106 years. Assuming that mice were infected with a single Sin Nombre virus genotype, phylogenetic analyses revealed that 10% (2/20) of viruses were reassortants, similar to the 14% (6/43) found in a previous report.

Conclusion

Age estimates from both segments suggest that Sin Nombre virus has evolved within the past 37–106 years. The rates of evolutionary changes reported here suggest that Sin Nombre virus M and S segment reassortment occurs frequently in nature.     

Species distribution models in climate change scenarios are still not useful for informing policy planning: an uncertainty assessment using fuzzy logic

We compared the effect of general circulation models and greenhouse gas emission scenarios on the uncertainty associated with models predicting changes in areas favourable to animal species. Given that mountain species are particularly at risk due to climate warming, we selected one amphibian (Baetic midwife toad), one reptile (Lataste's viper), one bird (Bonelli's eagle), and one mammal (Iberian wild goat) present in Spanish mountains to model their distributional response to climate change during this century. Climate forecasts for the whole century were provided by the Agencia Estatal de Meteorología (AEMET; National Meteorological Agency) of Spain, which adapted the general circulation models CGCM2 and ECHAM4 and produced expected temperature and precipitation values for Spain according to the A2 and B2 emission scenarios. We constructed separate models of the species response to spatial, topographic, human, and climate variables using current values of the corresponding variables. We predicted future areas favourable to the species by replacing the current climate values with those expected according to each climate change scenario, while keeping spatial, topographic and human variables constant. Fuzzy logic was used to compute the coincidence between predictions for different emission scenarios in the same global circulation model, and the consistency between predictions for the same emission scenario applying different general circulation models. In general, coincidences were higher than consistencies and, thus, discrepancies between predictions were more attributable to uncertainty in global circulation models, i.e. our insufficient knowledge concerning the effect of the oceans and atmosphere on climate, than to the putative effect of different emission scenarios on future climates. Our conclusion is that species distribution models in climate warming scenarios are still not useful for informing emission policy planning, although they have great potential as tools once consistencies become higher than coincidences.     

Atrial local Ca signaling and inositol 1,4,5-trisphosphate receptors

In atrial myocytes lacking t-tubules, action potential triggers junctional Ca²⁺ releases in the cell periphery, which propagates into the cell interior. The present article describes growing evidence on atrial local Ca²⁺ signaling and on the functions of inositol 1,4,5-trisphosphate receptors (IP₃Rs) in atrial myocytes, and show our new findings on the role of IP₃R subtype in the regulation of spontaneous focal Ca²⁺ releases in the compartmentalized areas of atrial myocytes. The Ca²⁺ sparks, representing focal Ca²⁺ releases from the sarcoplasmic reticulum (SR) through the ryanodine receptor (RyR) clusters, occur most frequently at the peripheral junctions in isolated resting atrial cells. The Ca²⁺ sparks that were darker and longer lasting than peripheral and non-junctional (central) sparks, were found at peri-nuclear sites in rat atrial myocytes. Peri-nuclear sparks occurred more frequently than central sparks. Atrial cells express larger amounts of IP₃Rs compared with ventricular cells and possess significant levels of type 1 IP₃R (IP₃R1) and type 2 IP₃R (IP₃R2). Over the last decade the roles of atrial IP₃R on the enhancement of Ca²⁺-induced Ca²⁺ release and arrhythmic Ca²⁺ releases under hormonal stimulations have been well documented. Using protein knock-down method and confocal Ca²⁺ imaging in conjunction with immunocytochemistry in the adult atrial cell line HL-1, we could demonstrate a role of IP₃R1 in the maintenance of peri-nuclear and non-junctional Ca²⁺ sparks via stimulating a posttranslational organization of RyR clusters.     

Predicting and Correcting Electrocution of Birds in Mediterranean Areas

ABSTRACT Bird electrocution on power lines is an important conservation problem that affects many endangered species. We surveyed 3,869 pylons in the Barcelona Pre-littoral Mountains (Catalonia, NE Spain) and collected 141 carcasses of electrocuted birds, mainly raptors and corvids. Univariate analysis indicated that metal pylons with pin-type insulators or exposed jumpers, with connector wires, located on ridges, overhanging other landscape elements, and in open habitats with low vegetation cover were the most dangerous. A logistic regression model indicated that the probability of a pylon electrocuting a bird was mainly related to pylon conductivity, distribution of the conductive elements on the cross-arms, cross-arm configuration, habitat, topography, whether the pylon was overhanging other landscape elements, and presence of rabbits (Oryctolagus cuniculus). We validated the predictive power of this model by using a random sample of 20% of all pylons surveyed. We found that bird mortality was aggregated mainly on pylons assigned a high probability risk by the model. Pylons included in the very high electrocution risk category (9.2%) accounted for 53.2% of carcasses, whereas pylons classified in the low electrocution risk category (54.5%) only accounted for 3.5% of mortality. Power companies employed this classification to prioritize the correction of 222 pylons by installing alternate cross-arms and suspended jumpers and isolating wires and jumpers. We evaluated the effectiveness of this mitigation strategy. A significant fall in the mortality rate on corrected pylons combined with the lack of any reduction in the mortality rate in a sample of 350 noncorrected pylons indicated that the model selected adequately the most dangerous pylons and that the applied correction measures were effective. Consequently, our strategy may be a useful tool for optimizing efforts and resources invested in solving the problem of bird electrocution.     

Optimal control of a rabies epidemic model with a birth pulse

A system of ordinary differential equations describes the population dynamics of a rabies epidemic in raccoons. The model accounts for the dynamics of a vaccine, including loss of vaccine due to animal consumption and loss from factors other than racoon uptake. A control method to reduce the spread of disease is introduced through temporal distribution of vaccine packets. This work incorporates the effect of the seasonal birth pulse in the racoon population and the attendant increase in new-borns which are susceptible to the diseases, analysing the impact of the timing and length of this pulse on the optimal distribution of vaccine packets. The optimization criterion is to minimize the number of infected raccoons while minimizing the cost of distributing the vaccine. Using an optimal control setting, numerical results illustrate strategies for distributing the vaccine depending on the timing of the infection outbreak with respect to the birth pulse.