An order-specific monoclonal antibody to Diptera reveals the impact of alternative prey on spider feeding behavior in a complex food web

Generalist predators have the capacity to restrict pest population growth, especially early in the season before densities increase. However, their polyphagous feeding habits sometimes translate into reduced pest consumption when they target alternative prey. An order-specific monoclonal antibody was developed to examine the strength of trophic connections between Diptera, a major category of non-pest prey, and linyphiid spiders in alfalfa. We report the development and characterization of a monoclonal antibody with order-level specificity to Diptera. This antibody elicited strong absorbance to 22 Diptera from 13 families, no false-positive reactivity to non-dipteran invertebrates, and antigen detection periods following prey consumption that were comparable between spiders. Over 900 field-collected females of the linyphiid spiders Erigone autumnalis and Bathyphantes pallidus were screened for Diptera antigen. Significantly more B. pallidus screened positive for Diptera (40%) compared to E. autumnalis (16%), indicating differential reliance on these prey. In parallel with the collection of spiders for gut-content analysis, prey availability was estimated at web sites. The two spiders exhibited different feeding responses to prey availability. Consumption of Diptera by B. pallidus was strongly correlated with Diptera abundance whilst the availability of other potential prey did not influence predation rates. Conversely, E. autumnalis did not prey upon Diptera in proportion to availability, but increased Collembola activity-density reduced dipteran consumption. Integration of molecular gut-content analysis with precise sampling of prey demonstrated how two closely related linyphiid spiders exhibit different feeding responses to the availability of prey under natural field conditions. Elucidating the feeding preferences of natural enemies is critical to effective incorporation of biological control by generalist predators in the management of agricultural pests.     

Selective inactivation of the Arg-Gly-Asp-Ser (RGDS) binding site in von Willebrand factor by site-directed mutagenesis.

In order to assess the requirement for the Arg-Gly-Asp-Ser (RGDS) consensus adhesion sequence in von Willebrand factor (vWF) for vWF binding to platelets and endothelial cells, point mutations were introduced into this sequence by site-directed mutagenesis. A glycine to alanine substitution yielded RADS-vWF, while an aspartate to glutamate substitution resulted in RGES-vWF. Recombinant RADS-vWF and RGES-vWF, purified from transformed Chinese hamster ovary cells, were compared with recombinant wild type vWF (WT-vWF) in functional assays with platelets and human umbilical vein endothelial cells (HU-VECs). High molecular weight RADS-vWF and RGES-vWF multimers inhibited binding of 125I-vWF to a mixture of insolubilized native type I and III collagen and competed effectively with 125I-vWF for binding to formalin-fixed platelets in the presence of ristocetin, indicating functional collagen and platelet glycoprotein Ib binding. However, RADS-vWF and RGES-vWF were unable to displace the binding of 125I-vWF to thrombin or ADP-activated platelets. The attachment of HUVECs to either RADS-vWF or RGES-vWF coated surfaces was reduced and spreading was almost completely inhibited, compared with WT-vWF. We conclude that point mutations of the RGDS sequence in vWF selectively impair binding to platelet glycoprotein IIb/IIIa and the HUVEC vitronectin receptor.     

Evaluation of past and potential phosphorus uptake at the Orlando Easterly Wetland

The Orlando Easterly Wetland (OEW), located near Christmas, Florida, USA, is among the longer-lived treatment wetlands in the United States. It was established in the late 1980s to reduce nitrogen and phosphorus concentrations from tertiary treated wastewater bound for the St. Johns River. A goal of 0.07 mg/l total phosphorus concentration has been set by the regulating agency (St. Johns River Water Management District). In order to understand and define the operating conditions for which this target could be met, a systematic study of historic phosphorus uptake was performed using a traditional first-order model. Phosphorus uptake performance is shown to correlate well with hydraulic performance for two parallel upstream cells. The first-order model is enhanced with predictive capabilities that acknowledge the correlation between the phosphorus uptake rate constant and the hydraulic loading rate observed in the system. Inherent limitations with the first-order modeling approach are addressed and uncertainty in model performance is used to bound predictions.     

Localized reversible frameshift mutation in an adhesin gene confers a phase-variable adherence phenotype in mycoplasma

The variable adherence-associated (Vaa) antigen of Mycoplasma hominis is an abundant surface lipoprotein adhesin that may mediate important interactions of this wall-less prokaryotic pathogen with the human host. Extensive mutational variation of Vaa size, as well as sequence and antigenic divergence, has been described previously. Using a series of clonal isolates representing an isogenic lineage of variants oscillating in Vaa expression, Vaa is further shown in this study to undergo high-frequency phase variation in expression, which correlated precisely with the ability of M . hominis to adhere to cultured human cells. Although no DNA rearrangements or sequence differences in the 5' regions flanking vaa alleles were detected between Vaa⁺ and Vaa⁻ variants, intragenic vaa sequences from this lineage revealed an oscillating mutation involving a single nucleotide deletion/insertion in a short tract of adenine residues near the 5' end of the mature Vaa coding sequence, which created a translational frameshift resulting in either a complete Vaa ORF or an in-frame UAG stop codon immediately downstream of the poly-A tract. Evidence for the occurrence of this high-frequency frameshift mutation in vivo was obtained from analysis of PCR-generated vaa sequences amplified from the joint synovial fluid of a patient with M . hominis -associated arthritis, which indicated that Vaa phase variation occurs during M . hominis infection in the natural host. These results identify a distinctive frameshift mutator element in the vaa gene that governs M . hominis adherence and highlight the importance of mutational alteration of primary gene products on the mycoplasma surface as a means of generating and maintaining functional diversity in the host.     

Paleolimnological evidence for recent acidification of Big Moose Lake,Adirondack Mountains,N.Y. (USA)

Big Moose L. has become significantly more acidic since the 1950s, based on paleolimnological analyses of sediment cores. Reconstruction of past lakewater pH using diatom assemblage data indicates that from prior to 1800 to ca. 1950, lakewater pH was about 5.8. After the mid-1950s, the inferred pH decreased steadily and relatively quickly to about 4.6. Alkalinity reconstructions indicate a decrease of about 30 eq · l^-1 during the same period. There was a major shift in diatom assemblage composition, including a nearly total loss of euplanktonic taxa. Chrysophyte scale assemblages and chironomid (midge larvae remains also changed in a pattern indicating decreasing lakewater pH starting in the 1950s. Accumulation rates of total Ca, exchangeable and oxide Al, and other metals suggest recent lake-watershed acidification. Cores were dated using²¹⁰Pb, pollen, and charcoal. Indicators of watershed change (deposition rates of Ti, Si, Al) do not suggest any major erosional events resulting from fires or logging. Accumulation rates of materials associated with combustion of fossil fuels (polycyclic aromatic hydrocarbons, coal and oil soot particles, some trace metals, and sulfur) are low until the late 1800s-early 1900s and increase relatively rapidly until the 1920s–1930s. Peak rates occurred between the late 1940s and about 1970, when rates declined.The recent decrease in pH of Big Moose L. cannot be accounted for by natural acidification or processes associated with watershed disturbance. The magnitude, rate and timing of the recent pH and alkalinity decreases, and their relationship to indicators of coal and oil combustion, indicate that the most reasonable explanation for the recent acidification is increased atmospheric deposition of strong acids derived from combustion of fossil fuels.     

Chilling-enhanced photooxidation : evidence for the role of singlet oxygen and superoxide in the breakdown of pigments and endogenous antioxidants

Chilling temperatures (5°C) and high irradiance (1000 microeinsteins per square meter per second) were used to induce photooxidation in detached leaves of cucumber (Cucumis sativus L.), a chilling-sensitive plant. Chlorophyll a, chlorophyll b, β carotene, and three xanthophylls were degraded in a light-dependent fashion at essentially the same rate. Lipid peroxidation (measured as ethane evolution) showed an O₂ dependency. The levels of three endogenous antioxidants, ascorbate, reduced glutathione, and α tocopherol, all showed an irradiance-dependent decline. α-Tocopherol was the first antioxidant affected and appeared to be the only antioxidant that could be implicated in long-term protection of the photosynthetic pigments. Results from the application of antioxidants having relative selectivity for ¹O₂, O₂⁻, or OH indicated that both ¹O₂ and O₂⁻ were involved in the chilling- and light-induced lipid peroxidation which accompanied photooxidation. Application of D₂O (which enhances the lifetime of ¹O₂) corroborated these results. Chilling under high light produced no evidence of photooxidative damage in detached leaves of chilling-resistant pea (Pisum sativum L.). Our results suggest a fundamental difference in the ability of pea to reduce the destructive effects of free-radical and ¹O₂ production in chloroplasts during chilling in high light.