Microhabitats, thermal heterogeneity, and patterns of physiological stress in the rocky intertidal zone.

Thermal stress has been considered to be among the most important determinants of organismal distribution in the rocky intertidal zone. Yet our understanding of how body temperatures experienced under field conditions vary in space and time, and of how these temperatures translate into physiological performance, is still rudimentary. We continuously monitored temperatures at a site in central California for a period of two years, using loggers designed to mimic the thermal characteristics of mussels, Mytilus californianus. Model mussel temperatures were recorded on both a horizontal and a vertical, north-facing microsite, and in an adjacent tidepool. We periodically measured levels of heat shock proteins (Hsp70), a measure of thermal stress, from mussels at each microsite. Mussel temperatures were consistently higher on the horizontal surface than on the vertical surface, and differences in body temperature between these sites were reflected in the amount of Hsp70. Seasonal peaks in extreme high temperatures ("acute" high temperatures) did not always coincide with peaks in average daily maxima ("chronic" high temperatures), suggesting that the time history of body temperature may be an important factor in determining levels of thermal stress. Temporal patterns in body temperature during low tide were decoupled from patterns in water temperature, suggesting that water temperature is an ineffective metric of thermal stress for intertidal organisms. This study demonstrates that spatial and temporal variability in thermal stress can be highly complex, and "snapshot" sampling of temperature and biochemical indices may not always be a reliable method for defining thermal stress at a site.     

Change and stabilization of the amyloid-beta(1-40) secondary structure by fluorocompounds

The misfolding of the amyloid peptide, which is the result of a well-known alpha-to-beta transition, causes neurodegenerative disorder. Fluorinated alcohols have been described in the literature as potent solvents which can refold the beta-conformation. The present studies demonstrate the effectiveness of differently fluorinated alcohols for the beta-to-alpha refolding process on fibrillar aggregated amyloid beta(1-40). The regenerated helical structure is shown to be maintained in the absence of the fluoroalcohols, a behaviour which was found to contrast with immunoglobulin. We interpret this difference on the basis of the hydrophilic/hydrophobic domains in the amyloid sequence and present some speculations regarding the free-energy levels of the folded states of both proteins. The effect of the -CF(3) group on the observed conformational changes is interpreted as a result of alterations of the hydration shell of the peptides. Moreover, based on the results achieved with fluoroalcohols, we have used novel fluorinated amphiphiles possessing blood-compatibility properties and studied their effect on amyloid beta(1-40). First results point in the direction of a beta-to-alpha transition. Therefore, the use of fluorine groups in the development of new drugs is considered a new possibility requiring further investigation for the prevention of amyloidosis.     

Physiological ecology of rocky intertidal organisms: a synergy of concepts

The rocky intertidal zone is among the most physically harshenvironments on earth. Marine invertebrates and algae livingin this habitat are alternatively pounded by waves and exposedto thermal extremes during low tide periods (Denny and Wethey,2001). Additionally, they must deal with strong selective pressuresrelated to predation and competition for space (Connell, 1961).As a result, the steep physical gradient and spatially condensedcommunity has made the rocky intertidal zone an ideal "naturallaboratory" to study the coupled role of physical and biologicalfactors in determining the abundance and distribution of organismsin nature (Connell, 1961; Paine, 1966, 1994).     

Thermal tolerance and climate warming sensitivity in tropical snails

Tropical ectotherms are predicted to be especially vulnerable to climate change because their thermal tolerance limits generally lie close to current maximum air temperatures. This prediction derives primarily from studies on insects and lizards and remains untested for other taxa with contrasting ecologies. We studied the HCT (heat coma temperatures) and ULT (upper lethal temperatures) of 40 species of tropical eulittoral snails (Littorinidae and Neritidae) inhabiting exposed rocky shores and shaded mangrove forests in Oceania, Africa, Asia and North America. We also estimated extremes in animal body temperature at each site using a simple heat budget model and historical (20 years) air temperature and solar radiation data. Phylogenetic analyses suggest that HCT and ULT exhibit limited adaptive variation across habitats (mangroves vs. rocky shores) or geographic locations despite their contrasting thermal regimes. Instead, the elevated heat tolerance of these species (HCT = 44.5 ± 1.8°C and ULT = 52.1 ± 2.2°C) seems to reflect the extreme temperature variability of intertidal systems. Sensitivity to climate warming, which was quantified as the difference between HCT or ULT and maximum body temperature, differed greatly between snails from sunny (rocky shore; Thermal Safety Margin, TSM = −14.8 ± 3.3°C and −6.2 ± 4.4°C for HCT and ULT, respectively) and shaded (mangrove) habitats (TSM = 5.1 ± 3.6°C and 12.5 ± 3.6°C). Negative TSMs in rocky shore animals suggest that mortality is likely ameliorated during extreme climatic events by behavioral thermoregulation. Given the low variability in heat tolerance across species, habitat and geographic location account for most of the variation in TSM and may adequately predict the vulnerability to climate change. These findings caution against generalizations on the impact of global warming across ectothermic taxa and highlight how the consideration of nonmodel animals, ecological transitions, and behavioral responses may alter predictions of studies that ignore these biological details.     

A Predominant Multidrug-Resistant Salmonella enterica Serovar Saintpaul Clonal Line in German Turkey and Related Food Products

Recently, Salmonella enterica subsp. enterica serovar Saintpaul has increasingly been observed in several countries, including Germany. However, the pathogenic potential and epidemiology of this serovar are not very well known. This study describes biological attributes of S. Saintpaul isolates obtained from turkeys in Germany based on characterization of their pheno- and genotypic properties. Fifty-five S. Saintpaul isolates from German turkeys and turkey-derived food products isolated from 2000 to 2007 were analyzed by using antimicrobial agent, organic solvent, and disinfectant susceptibility tests, isoelectric focusing, detection of resistance determinants, plasmid profiling, pulsed-field gel electrophoresis (PFGE), and hybridization experiments. These isolates were compared to an outgroup consisting of 24 S. Saintpaul isolates obtained from humans and chickens in Germany and from poultry and poultry products (including turkeys) in Netherlands. A common core resistance pattern was detected for 27 German turkey and turkey product isolates. This pattern included resistance (full or intermediate) to ampicillin, amoxicillin-clavulanic acid, gentamicin, kanamycin, nalidixic acid, streptomycin, spectinomycin, and sulfamethoxazole and intermediate resistance or decreased susceptibility to ciprofloxacin (MIC, 2 or 1 μg/ml, respectively) and several third-generation cephalosporins (including ceftiofur and cefoxitin [MIC, 4 to 2 and 16 to 2 μg/ml, respectively]). These isolates had the same core resistance genotype, with bla_TEM-1, aadB, aadA2, sul1, a Ser83→Glu83 mutation in the gyrA gene, and a chromosomal class 1 integron carrying the aadB-aadA2 gene cassette. Their XbaI, BlnI, and combined XbaI-BlnI PFGE patterns revealed levels of genetic similarity of 93, 75, and 90%, respectively. This study revealed that a multiresistant S. Saintpaul clonal line is widespread in turkeys and turkey products in Germany and was also detected among German human fecal and Dutch poultry isolates.Over the last few decades, the emergence and spread of antimicrobial agent-resistant zoonotic bacteria has become a serious public health concern (2, 23). The widespread use of antimicrobial agents for disease control, including at the farm level, has increased selection of antimicrobial agent-resistant Salmonella isolates (1, 23, 44). Food animals are considered an important reservoir for resistant bacteria. These animals and food products derived from them are traded worldwide, which contributes to the global spread of zoonotic agents and antimicrobial resistance. In the last few years, several monitoring activities were initiated in order to generate baseline data on antimicrobial resistance in bacteria isolated from livestock and food derived from animals that could be used in future assessments of the risk of antimicrobial resistance (10).According to European Union (EU) Zoonoses Regulation (EC) no. 2160/2003 on the control of Salmonella and other specified food-borne zoonotic agents, a European Community target for reducing the prevalence of Salmonella in turkey flocks had to be established. Consequently, EU Commission decision 2006/662/EC was released, and a baseline survey of the prevalence of Salmonella in turkey flocks was carried out in all European countries, including Germany, over a 1-year period starting on 1 October 2006 (http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178706574172.htm). The main objective of this study was to estimate the prevalence of Salmonella in commercial flocks of turkeys. The data showed that at the EU level Salmonella enterica serovar Bredeney was the serovar reported most frequently for fattening turkey flocks and occurred in 17.2% of the samples from Salmonella-positive flocks (1,084 of 3,702 flocks were positive), followed by S. enterica serovar Hadar, S. enterica serovar Derby, and then S. enterica serovar Saintpaul (14.0%, 11.3%, and 10.4% of the samples from positive flocks, respectively). In this study, S. Saintpaul was detected in fattening turkeys in 12 countries, reflecting the wide spread of this serovar. Recently, S. Saintpaul has been increasingly observed in several countries, including Germany. According to Enter-Net reports (data on Salmonella human isolates identified by European national reference centers), for the last quarter of the year 2006 S. Saintpaul was the fourth most common serovar (1.6%) and, in contrast to the data for previous years, was one of the most frequent causes of human salmonellosis in Europe. After this, its prevalence was 1.2% and 0.6% in the first quarters of 2007 and 2008, respectively, among the Salmonella serotypes implicated in human disease (http://ecdc.europa.eu/en/publications/Pages/Surveillance_Reports.aspx). During the period from 2001 to 2009 in Germany, 463 cases of human salmonellosis related to S. Saintpaul (0.09% of all cases; the maximum prevalence was 0.15% in 2008, the prevalence was 0.1% in 2002, 2005, 2006, and 2009 and 0.06% in 2004, and the minimum prevalence was 0.05% in 2007) were reported to the Robert Koch Institute (Berlin, Germany) (www3.rki.de/SurvStat). In Germany, S. Saintpaul attracted public attention particularly in 1993, when it caused a nationwide food-borne outbreak (27). This serotype has often been related to outbreaks in other countries, and in 2008 it was implicated in a large multistate human outbreak associated with various vegetables in the United States (4).Previous studies showed that isolates of S. Saintpaul are often multidrug resistant (33, 35), but little is known about the mechanisms underlying antimicrobial resistance or about the pathogenic potential and epidemiology of isolates belonging to this serotype. The goals of this study were to obtain information about the resistance characteristics of isolates collected between 2000 and 2007 in Germany and to assess possible clonal relationships. The isolates used originated from turkey feces collected during the German Salmonella baseline study (in 2006 and 2007) or from diagnostic samples, including samples of turkey feces and turkey-related food products. These isolates were compared with German strains isolated from humans and chickens and with poultry strains isolated in Netherlands.     

Dispersion of the bio-aerosol produced by the oak processionary moth

The oak processionary moth (Thaumetopoea processionea L.) is found in oak forests in most European countries. The caterpillars bear urticating hairs (setae) as a chemical defence. These hairs break off and are small enough to become airborne and be transported by the wind. Upon contact with humans the toxin can cause an allergic reaction that ranges from a skin rash to respiratory distress. In order to measure the terminal settling velocity of this bioaerosol, we used a small elutriator and tested its functionality with particles of known aerodynamic diameter. We determined that the mean settling velocity of the setae is about 1 cm/s, corresponding to an aerodynamic diameter of 19 μm for setae with a diameter of 6 μm and a length of 190 μm. The dispersion of the hairs in the atmosphere for a typical summer day was calculated by means of an Eulerian model. The results of this calculation revealed that the maximum concentrations in the atmosphere on a typical summer day reach 20–30% of the concentration found directly at the source. Those maximum concentrations are reached at a distance from the source that varies between 174 and 562 m, depending on the atmospheric stability and the settling velocity.

Simpler is better: fewer non‐target insects trapped with a four‐component chemical lure vs. a chemically more complex food‐type bait for Drosophila suzukii

Baits – fermented food products – are generally attractive to many types of insects, which makes it difficult to sort through non‐target insects to monitor a pest species of interest. We test the hypothesis that a chemically simpler and more defined attractant developed for a target insect is more specific and attracts fewer non‐target insects than a chemically more complex food‐type bait. A four‐component chemical lure isolated from a food bait and optimized for the spotted wing drosophila (SWD), Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), was compared to the original wine/vinegar bait to assess the relative responses of non‐target insects. In several field experiments in Washington State, USA, it was shown that numbers of pest muscid flies, cutworm and armyworm moths, and pest yellowjackets were reduced in traps baited with the chemical lure compared to the wine/vinegar bait. In other field experiments in the states of Washington, Oregon, and New York, numbers of non‐target drosophilid flies were also reduced in traps baited with the chemical lure relative to wine/vinegar bait. In Washington, numbers of Drosophila melanogaster Meigen and Drosophila obscura Fallen species groups and Drosophila immigrans Sturtevant were reduced in the chemical lure traps, whereas in New York, D. melanogaster and D. obscura species groups, D. immigrans, Drosophila putrida Sturtevant, Drosophila simulans Sturtevant, Drosophila tripunctata Loew, and Chymomyza spp. numbers were reduced. In Oregon, this same effect was observed with the D. melanogaster species group. Taken together, these results indicate that the four‐component SWD chemical lure will be more selective for SWD compared to fermentation baits, which should reduce time and cost involved in trapping in order to monitor SWD.     

Warfarin anticoagulation exacerbates the risk of hemorrhagic transformation after rt-PA treatment in experimental stroke: therapeutic potential of PCC

Background

Oral anticoagulant therapy (OAT) with warfarin is the standard of stroke prevention in patients with atrial fibrillation. Approximately 30% of patients with cardioembolic strokes are on OAT at the time of symptom onset. We investigated whether warfarin exacerbates the risk of thrombolysis-associated hemorrhagic transformation (HT) in a mouse model of ischemic stroke.

Methods

62 C57BL/6 mice were used for this study. To achieve effective anticoagulation, warfarin was administered orally. We performed right middle cerebral artery occlusion (MCAO) for 3 h and assessed functional deficit and HT blood volume after 24 h.

Results

In non-anticoagulated mice, treatment with rt-PA (10 mg/kg i.v.) after 3 h MCAO led to a 5-fold higher degree of HT compared to vehicle-treated controls (4.0±0.5 µl vs. 0.8±0.1, p<0.001). Mice on warfarin revealed larger amounts of HT after rt-PA treatment in comparison to non-anticoagulated mice (9.2±3.2 µl vs. 2.8±1.0, p<0.05). The rapid reversal of anticoagulation by means of prothrombin complex concentrates (PCC, 100 IU/kg) at the end of the 3 h MCAO period, but prior to rt-PA administration, neutralized the exacerbated risk of HT as compared to sham-treated controls (3.8±0.7 µl vs. 15.0±3.8, p<0.001).

Conclusion

In view of the vastly increased risk of HT, it seems to be justified to withhold tPA therapy in effectively anticoagulated patients with acute ischemic stroke. The rapid reversal of anticoagulation with PCC prior to tPA application reduces the risk attributed to warfarin pretreatment and may constitute an interesting therapeutic option.     

Evolution and population structure of Salmonella enterica serovar Newport

Salmonellosis caused by Salmonella enterica serovar Newport is a major global public health concern, particularly because S. Newport isolates that are resistant to multiple drugs (MDR), including third-generation cephalosporins (MDR-AmpC phenotype), have been commonly isolated from food animals. We analyzed 384 S. Newport isolates from various sources by a multilocus sequence typing (MLST) scheme to study the evolution and population structure of the serovar. These were compared to the population structure of S. enterica serovars Enteritidis, Kentucky, Paratyphi B, and Typhimurium. Our S. Newport collection fell into three lineages, Newport-I, Newport-II, and Newport-III, each of which contained multiple sequence types (STs). Newport-I has only a few STs, unlike Newport-II or Newport-III, and has possibly emerged recently. Newport-I is more prevalent among humans in Europe than in North America, whereas Newport-II is preferentially associated with animals. Two STs of Newport-II encompassed all MDR-AmpC isolates, suggesting recent global spread after the acquisition of the bla(CMY-2) gene. In contrast, most Newport-III isolates were from humans in North America and were pansusceptible to antibiotics. Newport was intermediate in population structure to the other serovars, which varied from a single monophyletic lineage in S. Enteritidis or S. Typhimurium to four discrete lineages within S. Paratyphi B. Both mutation and homologous recombination are responsible for diversification within each of these lineages, but the relative frequencies differed with the lineage. We conclude that serovars of S. enterica provide a variety of different population structures.