Aedes aegypti Mosquitoes Exhibit Decreased Repellency by DEET following Previous Exposure

DEET (N,N-Diethyl-m-toluamide) is one of the most widely used mosquito repellents. Although DEET has been shown to be extremely effective, recent studies have revealed that certain individual insects are unaffected by its presence. A genetic basis for this has been shown in Aedes aegypti mosquitoes and the fruit fly Drosophila melanogaster, but, for the triatomine bug, Rhodnius prolixus, a decrease in response to DEET occurred shortly after previous exposure, indicating that non-genetic factors may also be involved in DEET “insensitivity”. In this study, we examined host-seeking behaviour and electrophysiological responses of A. aegypti after pre-exposure to DEET. We found that three hours after pre-exposure the mosquitoes showed behavioural insensitivity, and electroantennography revealed this correlated with the olfactory receptor neurons responding less to DEET. The change in behaviour as a result of pre-exposure to DEET has implications for the use of repellents and the ability of mosquitoes to overcome them.     

Plant response to climate change along the forest‐tundra ecotone in northeastern Siberia

Russia's boreal (taiga) biome will likely contract sharply and shift northward in response to 21st century climatic change, yet few studies have examined plant response to climatic variability along the northern margin. We quantified climate dynamics, trends in plant growth, and growth–climate relationships across the tundra shrublands and Cajander larch (Larix cajanderi Mayr.) woodlands of the Kolyma river basin (657 000 km²) in northeastern Siberia using satellite‐derived normalized difference vegetation indices (NDVI), tree ring‐width measurements, and climate data. Mean summer temperatures (T_s) increased 1.0 °C from 1938 to 2009, though there was no trend (P > 0.05) in growing year precipitation or climate moisture index (CMI_gy). Mean summer NDVI (NDVI_s) increased significantly from 1982 to 2010 across 20% of the watershed, primarily in cold, shrub‐dominated areas. NDVI_s positively correlated (P < 0.05) with T_s across 56% of the watershed (r = 0.52 ± 0.09, mean ± SD), principally in cold areas, and with CMI_gy across 9% of the watershed (r = 0.45 ± 0.06), largely in warm areas. Larch ring‐width measurements from nine sites revealed that year‐to‐year (i.e., high‐frequency) variation in growth positively correlated (P < 0.05) with June temperature (r = 0.40) and prior summer CMI (r = 0.40) from 1938 to 2007. An unexplained multi‐decadal (i.e., low‐frequency) decline in annual basal area increment (BAI) occurred following the mid‐20th century, but over the NDVI record there was no trend in mean BAI (P > 0.05), which significantly correlated with NDVI_s (r = 0.44, P < 0.05, 1982–2007). Both satellite and tree‐ring analyses indicated that plant growth was constrained by both low temperatures and limited moisture availability and, furthermore, that warming enhanced growth. Impacts of future climatic change on forests near treeline in Arctic Russia will likely be influenced by shifts in both temperature and moisture, which implies that projections of future forest distribution and productivity in this area should take into account the interactions of energy and moisture limitations.     

Direct observation of phytoplankton, TEP and aggregates on polycarbonate filters using brightfield microscopy

There has been little use of standard (i.e. non-inverted) microscopesfor observing and counting phytoplankton in filtered water samplesusing brightfield white light illumination due to light interferencefrom the filters. If filters are placed on newly designed frostedslides (Cyto-clear, Poretics Corp.), however, phytoplanktoncan be viewed directly on the surfaces of polycarbonate filtersunder brightfield illumination. Lake and seawater samples wereused to show that samples stained with alcian blue (to identifythe presence of paniculate polysaccharides) and analyzed withwhite light can also be simultaneously stained with fluorochromes(i.e. DAPI and acridine orange) for additional examination ofthe sample using fluorescent techniques. Black filters, whichare necessary for epifluorescent techniques, did not interferewith brightfield viewing. Using double staining techniques,we found that transparent exopolymer particles (TEP) recentlydiscovered in marine systems are also present in lakes. Notall aggregates in the fresh and seawater systems absorbed thealcian blue stain, however, indicating that not all amorphousparticles in these systems are rich in negatively charged polysaccharides.     

Linear skeletal extension rates in two species of Diploria from high-latitude reefs in Bermuda

X-radiography was used to study annual linear skeletal extension rates of the reef-building scleractinian corals Diploria strigosa and Diploria labyrinthiformis from the high-latitude reefs of Bermuda. Coral samples for X-radiography were collected from seven localities of varying biotopes and depths around the Bermuda platform and band couplets were measured. Mean extension rates of both species were highest on inshore and nearshore reefs, gradually decreasing towards the edge of the Bermuda platform and onto the fore-reef slope. Extension rates of D. labyrinthiformis were statistically higher than those of D. strigosa at three localities, while at the other four, the rates of both species were not statistically separable. extension rates of d. labyrinthiformis were statistically higher than D. strigosa within depths of 20 m and 32 m but not statistically separable at 3 m and 6 m depths. Extension rates of both species decreased significantly with increasing depth (r ²=0.92, P<0.03 for D. labyrinthiformis and r ²=0.95, P<0.02 for D. strigosa). Each species showed an inverse curvilinear relationship between extension rate and depth, the rate of change (i.e. slope) being the same for each species. Comparison of extension rates of each species from Berumuda with published rates of these species from lower latitudes showed an inverse relationship between extension rate and latitude.     

Insight into the Role of Substrate-binding Residues in Conferring Substrate Specificity for the Multifunctional Polysaccharide Lyase Smlt1473

Anionic polysaccharides are of growing interest in the biotechnology industry due to their potential pharmaceutical applications in drug delivery and wound treatment. Chemical composition and polymer length strongly influence the physical and biological properties of the polysaccharide and thus its potential industrial and medical applications. One promising approach to determining monomer composition and controlling the degree of polymerization involves the use of polysaccharide lyases, which catalyze the depolymerization of anionic polysaccharides via a β-elimination mechanism. Utilization of these enzymes for the production of custom-made oligosaccharides requires a high degree of control over substrate specificity. Previously, we characterized a polysaccharide lyase (Smlt1473) from Stenotrophomonas maltophilia k279a, which exhibited significant activity against hyaluronan (HA), poly-β-d-glucuronic acid (poly-GlcUA), and poly-β-d-mannuronic acid (poly-ManA) in a pH-regulated manner. Here, we utilize a sequence structure guided approach based on a homology model of Smlt1473 to identify nine putative substrate-binding residues and examine their effect on substrate specificity via site-directed mutagenesis. Interestingly, single point mutations H221F and R312L resulted in increased activity and specificity toward poly-ManA and poly-GlcUA, respectively. Furthermore, a W171A mutant nearly eliminated HA activity, while increasing poly-ManA and poly-GlcUA activity by at least 35%. The effect of these mutations was analyzed by comparison with the high resolution structure of Sphingomonas sp. A1-III alginate lyase in complex with poly-ManA tetrasaccharide and by taking into account the structural differences between HA, poly-GlcUA, and poly-ManA. Overall, our results demonstrate that even minor changes in active site architecture have a significant effect on the substrate specificity of Smlt1473, whose structural plasticity could be applied to the design of highly active and specific polysaccharide lyases.     

Do Shade-Grown Coffee Plantations Pose a Disease Risk for Wild Birds?

Shade-grown coffee plantations are often promoted as a conservation strategy for wild birds. However, these agro-ecosystems are actively managed for food production, which may alter bird behaviors or interactions that could change bird health, compared to natural forest. To examine whether there is a difference between the health parameters of wild birds inhabiting shade-grown coffee plantations and natural forest, we evaluated birds in Costa Rica for (1) their general body condition, (2) antibodies to pathogens, (paramyxovirus and Mycoplasma spp.), and (3) the prevalence and diversity of endo-, ecto-, and hemoparasites. We measured exposure to Mycoplasma spp. and paramyxovirus because these are pathogens that could have been introduced with domestic poultry, one mechanism by which these landscapes could be detrimental to wild birds. We captured 1,561 birds representing 75 species. Although seasonal factors influenced body condition, we did not find bird general body condition to be different. A total of 556 birds of 31 species were tested for antibodies against paramyxovirus-1. Of these, five birds tested positive, four of which were from shade coffee. Out of 461 other tests for pathogens (for antibodies and nucleotide detection), none were positive. Pterolichus obtusus, the feather mite of chickens, was found on 15 birds representing two species and all were from shade-coffee plantations. Larvated eggs of Syngamus trachea, a nematode typically associated with chickens, were found in four birds captured in shade coffee and one captured in forest. For hemoparasites, a total of 1,121 blood smears from 68 bird species were examined, and only one species showed a higher prevalence of infection in shade coffee. Our results indicate that shade-coffee plantations do not pose a significant health risk to forest birds, but at least two groups of pathogens may deserve further attention: Haemoproteus spp. and the diversity and identity of endoparasites.     

Dynamic energy budget models with size-dependent hazard rates

We formulate and analyze two dynamic energy budget models, a net assimilation model with constant allocation strategy and a net production model with a 2-stage allocation strategy, with the objective of determining strategies that maximize the expected lifetime reproductive energy. The per capita death rate depends on the organisms size, as for example when the main cause of death is predation. In the analysis of the net production model, the size at maturity is calculated along with the probability of reaching that size. We show that a small probability of survival to maturity is incompatible with the simple assumption of an exponential survival probability. We demonstrate that when the hazard rate is significantly greater for small individuals than large ones, it is possible for the optimum strategy to be for an individual to grow to a large size in spite of an arbitrarily small probability of survival to maturity. Numerical simulations indicate how the optimal allocation strategies depend on the parameter values.     

Association of the calpain/calpastatin network with subcellular organelles

The calcium-activated cysteine protease calpain is intimately involved in modulating cell adhesion and migration. The two ubiquitous isoforms of this protease, calpain I and II, are considered to be cytosolic proteins that can translocate to both focal complexes/adhesions or the plasma membrane. Using confocal microscopy and isopycnic density centrifugation, the results demonstrate that calpain I and II, the 30kDa regulatory subunit, and calpastatin associate with the endoplasmic reticulum and Golgi apparatus. Confocal microscopy reveals that calpain II colocalizes with the subcellular proteins calnexin and Rab6 in cells bound to laminin. To further verify this association, cell lysates prepared from laminin stimulated and unstimulated cells were subjected to isopycnic density centrifugation. The results reveal an increased association of calpain I, II, calpastatin, and the 30kDa regulatory subunit with the endoplasmic reticulum and Golgi apparatus as evidenced by their position in the gradient relative to calnexin, Rab6, caveolin, and beta1 integrin after laminin stimulation. This correlates with the accumulation of inducible calpain activity at the endoplasmic reticulum-Golgi apparatus interface. Further experiments established that calpain II colocalizes with phosphatidylinositol 4,5-bisphosphate. Finally, calpain II associates with membrane lipid rafts. These results provide new insights into how the calpain/calpastatin network is spatially and temporally regulated in cells binding to the extracellular matrix.     

Location,time, and temperature dependence of digestion in simple animal tracts

In this paper, we develop plug flow reactor models that simultaneously investigate how reaction and absorption, morphological differences, and temperature influence nutrient acquisition rates in simple, tubular animal guts. We present analytical solutions to the resulting reaction-advection equations that model these processes, and we obtain formulas giving the throughflow speed that maximizes the absorption rate. The model predicts that the optimal digestion speed increases as the ratio of the rate of enzyme breakdown to the rate of absorption increases.