Climate and on-farm risk factors associated with Giardia duodenalis cysts in storm runoff from California coastal dairies

Climatic factors and on-farm management practices were evaluated for their association with the concentrations (cyst/liter) and instantaneous loads (cysts/second) of Giardia duodenalis in storm-based runoff from dairy lots and other high-cattle-use areas on five coastal California farms over two storm seasons. Direct fluorescent antibody analysis was used to quantitate cysts in 350 storm runoff samples. G. duodenalis was detected on all five dairy farms, with fluxes of 1 to 14,000 cysts/liter observed in 16% of samples. Cysts were detected in 41% of runoff samples collected near cattle less than 2 months old, compared to 10% of runoff samples collected near cattle over 6 months old. Furthermore, the concentrations and instantaneous loads of cysts were > or =65 and > or =79 times greater, respectively, in runoff from sites housing young calves than in sites housing other age classes of animals. Factors associated with environmental loading of G. duodenalis included cattle age, cattle stocking number, and precipitation but not lot area, land slope, or cattle density. Vegetated buffer strips were found to significantly reduce waterborne cysts in storm runoff: each additional meter of vegetated buffer placed below high-cattle-use areas was associated with reductions in the concentration and instantaneous load of cysts by factors of 0.86 and 0.79 (-0.07 and -0.10 log(10)/m), respectively. Straw mulch, seed application, scraping of manure, and cattle exclusion did not significantly affect the concentration or load of G. duodenalis cysts. The study findings suggest that vegetated buffer strips, especially when placed near dairy calf areas, should help reduce the environmental loading of these fecal protozoa discharging from dairy farms.     

Diversity of ndo genes in mangrove sediments exposed to different sources of polycyclic aromatic hydrocarbon pollution

Polycyclic aromatic hydrocarbon (PAH) pollutants originating from oil spills and wood and fuel combustion are pollutants which are among the major threats to mangrove ecosystems. In this study, the composition and relative abundance in the sediment bacterial communities of naphthalene dioxygenase (ndo) genes which are important for bacterial adaptation to environmental PAH contamination were investigated. Three urban mangrove sites which had characteristic compositions and levels of PAH compounds in the sediments were selected. The diversity and relative abundance of ndo genes in total community DNA were assessed by a newly developed ndo denaturing gradient gel electrophoresis (DGGE) approach and by PCR amplification with primers targeting ndo genes with subsequent Southern blot hybridization analyses. Bacterial populations inhabiting sediments of urban mangroves under the impact of different sources of PAH contamination harbor distinct ndo genotypes. Sequencing of cloned ndo amplicons comigrating with dominant DGGE bands revealed new ndo genotypes. PCR-Southern blot analysis and ndo DGGE showed that the frequently studied nah and phn genotypes were not detected as dominant ndo types in the mangrove sediments. However, ndo genotypes related to nagAc-like genes were detected, but only in oil-contaminated mangrove sediments. The long-term impact of PAH contamination, together with the specific environmental conditions at each site, may have affected the abundance and diversity of ndo genes in sediments of urban mangroves.     

Biophysical properties and functional significance of stem water storage tissues in Neotropical savanna trees

Biophysical characteristics of sapwood and outer parenchyma water storage compartments were studied in stems of eight dominant Brazilian Cerrado tree species to assess the impact of differences in tissue capacitance on whole-plant water relations. The rate of decline in tissue water potential with relative water content (RWC) was greater in the outer parenchyma than in the sapwood for most of the species, resulting in tissue-and species-specific differences in capacitance. Sapwood capacitance on a tissue volume basis ranged from 40 to 160 kg m-3 MPa-1, whereas outer parenchyma capacitance ranged from 25 to only 60 kg m-3 MPa-1. In addition, osmotic potentials at full turgor and at the turgor loss point were more negative for the outer parenchyma compared with the sapwood, and the maximum bulk elastic modulus was higher for the outer parenchyma than for the sapwood. Sapwood capacitance decreased linearly with increasing sapwood density across species, but there was no significant correlation between outer parenchyma capacitance and tissue density. Midday leaf water potential, the total hydraulic conductance of the soil/leaf pathway and stomatal conductance to water vapour (gs) all increased with stem volumetric capacitance, or with the relative contribution of stored water to total daily transpiration. However, the difference between the pre-dawn water potential of non-transpiring leaves and the weighted average soil water potential, a measure of the water potential disequilibrium between the plant and soil, increased asymptotically with total stem capacitance across species, implying that overnight recharge of water storage compartments was incomplete in species with greater capacitance. Overall, stem capacitance contributes to homeostasis in the diurnal and seasonal water balance of Cerrado trees.     

Conformations generated during turnover of the Azotobacter vinelandii nitrogenase MoFe protein and their relationship to physiological function

Various S=3/2 EPR signals elicited from wild-type and variant Azotobacter vinelandii nitrogenase MoFe proteins appear to reflect different conformations assumed by the FeMo-cofactor with different protonation states. To determine whether these presumed changes in protonation and conformation reflect catalytic capacity, the responses (particularly to changes in electron flux) of the alphaH195Q, alphaH195N, and alphaQ191K variant MoFe proteins (where His at position 195 in the alpha subunit is replaced by Gln/Asn or Gln at position alpha-191 by Lys), which have strikingly different substrate-reduction properties, were studied by stopped-flow or rapid-freeze techniques. Rapid-freeze EPR at low electron flux (at 3-fold molar excess of wild-type Fe protein) elicited two transient FeMo-cofactor-based EPR signals within 1 s of initiating turnover under N(2) with the alphaH195Q and alphaH195N variants, but not with the alphaQ191K variant. No EPR signals attributable to P cluster oxidation were observed for any of the variants under these conditions. Furthermore, during turnover at low electron flux with the wild-type, alphaH195Q or alphaH195N MoFe protein, the longer-time 430-nm absorbance increase, which likely reflects P cluster oxidation, was also not observed (by stopped-flow spectrophotometry); it did, however, occur for all three MoFe proteins under higher electron flux. No 430-nm absorbance increase occurred with the alphaQ191K variant, not even at higher electron flux. This putative lack of involvement of the P cluster in electron transfer at low electron flux was confirmed by rapid-freeze (57)Fe M?ssbauer spectroscopy, which clearly showed FeMo-factor reduction without P cluster oxidation. Because the wild-type, alphaH195Q and alphaH195N MoFe proteins can bind N(2), but alphaQ195K cannot, these results suggest that P cluster oxidation occurs only under high electron flux as required for N(2) reduction.     

Surface potential determination in planar lipid bilayers: a simplification of the conductance-ratio method

One of the methods available for the measurement of surface potentials of planar lipid bilayers uses the conductance ratio between a charged and a neutral bilayer doped with ionophores to calculate the surface potential of the charged bilayer. We have devised a simplification of that method which does not require the use of an electrically neutral bilayer as control. The conductance of the charged bilayer is measured before and after the addition of divalent cations (Ba(2+)) to the bathing solution. Ba(2+) ions screen fixed surface charges, decreasing the surface potential. If the membrane is negatively charged the screening has the effect of decreasing the membrane conductance to cations. The resulting conductance ratio is used to calculate the surface potential change, which is fed into an iterative computer program. The program generates pairs of surface potential values and calculates the surface charge density for the two conditions. Since the surface charge density remains constant during this procedure, there is only one pair of surface potentials that satisfies the condition of constant charge density. Applying this method to experimental data from McLaughlin et al. [McLaughlin, S.G.A., Szabo, G. and Eisenman, G., Divalent ions and the surface potential of charged phospholipid membranes, J. Gen. Physiol., 58 (1971) 667-687.] we have found very similar results. We have also successfully used this method to determine the effect of palmitic acid on the surface potential of asolectin membranes.     

Cougar Predation on Black-and-Gold Howlers on Mutum Island,Southern Brazil<Superscript>1</Superscript>

Researchers consider predation rates by terrestrial animals to be lower in the case of arboreal primates, particularly among large-bodied species. We recorded the consumption of black-and-gold howlers (Alouatta caraya) by cougars (Puma concolor) as evidence of predation on an island of the upper Paraná River. We collected and processed fecal samples of the felid in 2004 and 2005. We identified items in the laboratory by comparison with museum specimens. We considered each species in a fecal sample as a single occurrence. Based on analysis of the cuticle scale pattern, we identified the felid as cougar. Howlers occurred in 4 out of the 8 fecal samples (40% of the occurrences). In addition to howlers, we also recorded 5 occurrences of agouti (Dasyprocta azarae; 50%) and a small unidentified sigmodontine rodent (10%). The abundance of howlers and the low forest canopy in a successional vegetation might have facilitated the predation of the large primates by a primarily terrestrial predator. The versatility of cougars is corroborated by the consumption of prey species that were abundant in the region and that were available in different forest strata, such as howlers and agoutis.     

Validation of a Plasmodium falciparum parasite transformed with green fluorescent protein for antimalarial drug screening

Aiming to replace the radioisotopic assay, the widely used procedure for vitro antimalarial drug screening, we set up a protocol using a Plasmodium falciparum strain transformed with the green fluorescent protein (PfGFP), which can be quickly and specifically quantified by flow cytometry. On the basis of a side-by-side comparison, this PfGFP-based method showed results similar to those obtained with the standard radioisotopic method.     

Effects of relative humidity on development, fecundity and survival of three storage mites

The developmental rate of immature stages and the reproduction of adults of Tyrophagus putrescentiae (Schrank), T. neiswanderi Johnston and Bruce and Acarus farris (Oudemans) were examined at 70, 80 and 90% r.h. and a constant temperature of 25°C. At 70% r.h., T. putrescentiae and A. farris immature stages failed to reach the protonymph stage as 100% of the larvae died, whereas T. neiswanderi was able to complete development. The developmental time of all immature stages for the three species was significantly increased as relative humidity was reduced. The mobile stages were particularly susceptible, as the time needed to complete their development at lower relative humidities suffered greater increases than the egg stage. At 70% r.h., T. putrescentiae and A. farris were not able to lay eggs and only 24% of T. neiswanderi pairs were fertile. The reproductive parameters of the three species at the relative humidities at which they were able to lay eggs showed significant differences, except for the percentage of fertile mating at 80 and 90% r.h. As relative humidity increased, preoviposition period was reduced and fecundity and daily fecundity was increased, whereas the oviposition period showed different patterns for the three species. The intrinsic rate of increase (r _m ) of T. neiswanderi at 70% r.h. was negative indicating that, at these conditions, mite populations of this species will diminish until they disappear. As relative humidity increased from 80 to 90% r.h. this parameter was almost twofold for both Tyrophagus species. The r _m obtained for A. farris at 90% r.h. was similar to that of T. neiswanderi at the same humidity while at 80% r.h. it was very small so that the population doubling time was more than 84 days. The influence of relative humidity on biology of these mites and its practical application as control measure are discussed.     

Regional ecosystem structure and function: ecological insights from remote sensing of tropical forests

Ecological studies in tropical forests have long been plagued by difficulties associated with sampling the crowns of large canopy trees and large inaccessible regions, such as the Amazon basin. Recent advances in remote sensing have overcome some of these obstacles, enabling progress towards tackling difficult ecological problems. Breakthroughs have helped transform the dialog between ecology and remote sensing, generating new regional perspectives on key environmental gradients and species assemblages with ecologically relevant measures such as canopy nutrient and moisture content, crown area, leaf-level drought responses, woody tissue and surface litter abundance, phenological patterns, and land-cover transitions. Issues that we address here include forest response to altered precipitation regimes, regional disturbance and land-use patterns, invasive species and landscape carbon balance.