Cation exchange capacity of density fractions from paired conifer/grassland soils

The cation exchange capacity (CEC) of a soil depends on the type and amount of both mineral and organic surfaces. Previous studies that have sought to determine the relative contribution of organic matter to total soil CEC have not addressed differences in soil organic matter (SOM) composition that could lead to differences in CEC. The objectives of this study were (1) to compare the CEC of two distinct SOM pools, the “light fraction (LF)” composed of particulate plant, animal, and microbial debris, and the “heavy fraction (HF)” composed of mineral-bound organic matter; and (2) to examine the effects of differences in aboveground vegetation on CEC. Soil samples were collected from four paired grassland/conifer sites within a single forested area and density fractionated. LF CEC was higher in conifer soils than in grassland soils, but there was no evidence of an effect of vegetation on CEC for the HF or bulk soil. LF CEC (but not HF CEC) correlated well with the C concentration in the fraction. The mean CEC of both fractions (per kg fraction) exceeded that of the bulk soil; thus, when the LF and HF CEC were combined mathematically by weighting values for each fraction in proportion to dry mass, the resulting value was nearly twice the measured CEC of bulk soil. On a whole soil basis, the HF contributed on average 97% of the CEC of the whole soil, although this conclusion must be tempered given the inflation of CEC values by the density fractionation procedure.     

Glycoprofiling of bifidobacterial consumption of human milk oligosaccharides demonstrates strain specific, preferential consumption of small chain glycans secreted in early human lactation

The molecular basis by which human breast milk supports the development of a protective intestinal microbiome in infants is unknown. After lactose and lipids, human milk oligosaccharides (HMOs) are quantitatively the third largest and most diverse component of breast milk. In this work, glycomic profiling of HMO consumption by bifidobacteria using Fourier transform ion cyclotron resonance mass spectrometry reveals that one species, Bifidobacterium longum biovar infantis ATCC 15697, an isolate from the infant gut, preferentially consumes small mass oligosaccharides, representing 63.9% of the total HMOs available. These HMOs were detected in human breast milk at the onset and constantly through the first month of lactation by use of high performance liquid chromatography-chip time-of-flight mass spectrometry. Further characterization revealed that strain ATCC 15697 possesses both fucosidase and sialidase activities not present in the other tested strains. This work provides evidence that these small mass HMOs are selectively metabolized by select bifidobacterial strains and represent a potential new class of bioactive molecules functioning as prebiotics to facilitate a protective gut colonization in breast-fed newborns.     

Low α-Amylase Starch Digestibility of Cooked Sorghum Flours and the Effect of Protein

The comparably low starch digestibility of cooked sorghum flours was studied with reference to normal maize. Four sorghum cultivars that represent different types of endosperm were used. Starch digestibilities of 4% cooked sorghum flour suspensions, measured as reducing sugars liberated following α-amylase digestion, were 15–25% lower than for cooked maize flour, but there were no differences among the cooked pure starches. After the flours were predigested with pepsin to remove some proteins, the starch digestibility of cooked sorghum flours increased 7–14%, while there was only 2% increase in normal maize; however, there was no effect of pepsin treatment on starch digestibility if the flours were first cooked and then digested. After cooking with reducing agent, 100 mM sodium metabisulfite, starch digestibility of sorghum flours increased significantly while no significant effect was observed for maize. Also, starch solubility of sorghum flours at 85 and 100°C was lower than in maize, and sodium metabisulfite increased solubility much more in sorghum than in maize. Differential scanning calorimetry results of the flour residue after α-amylase digestion did not show any peaks over a temperature range of 20–120°C, indicating that sorghum starches had all undergone gelatinization. These findings indicate that the protein in cooked sorghum flour pastes plays an important role in making a slowly digesting starch.     

Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

N2O, NO, and NH3 Emissions from Soil after the Application of Organic Fertilizers, Urea and Water

Agricultural soil is a major source of nitrous oxide (N₂O), nitric oxide (NO) and ammonia (NH₃). Little information is available on emissions of these gases from soils amended with organic fertilizers at different soil water contents. N₂O, NO and NH₃ emissions were measured in large-scale incubations of a fresh sandy loam soil and amended with four organic fertilizers, [poultry litter (PL), composted plant residues (CP), sewage sludge pellets (SP) and cattle farm yard manure (CM)], urea fertilizer (UA) or a zero-N control (ZR) for 38 days. Fertilizers were added to soil at 40, 60 or 80% water-filled pore space (WFPS). The results showed that urea and organic fertilizer were important sources of N₂O and NO. Total N₂O and NO emissions from UA ranged from 0.04 to 0.62%, and 0.23 to 1.55% of applied N, respectively. Total N₂O and NO emissions from organic fertilizer treatments ranged from 0.01 to 1.65%, and <0.01 to=" 0.55%=" of=" applied=" n,=" respectively.=" the=" lower=">₂O and NO emissions from CP and CM suggested that applying N is these forms could be a useful mitigation option. Comparison of the NO-N/N₂O-N ratio suggested that nitrification was more dominant in UA whereas denitrification was more dominant in the organic fertilizer treatments. Most N was lost from PL and UA as NH₃, and this was not influenced significantly by WFPS. Emissions of NH₃ from UA and PL ranged from 62.4 to 69.6%, and 3.17 to 6.11% of applied N, respectively.     

Using Epiphytic Macrolichen Communities for Biomonitoring Ammonia in Forests of the Greater Sierra Nevada, California

Chronic, excessive nitrogen deposition is potentially an important ecological threat to forests of the greater Sierra Nevada in California. We developed a model for ammonia bioindication, a major nitrogen pollutant in the region, using epiphytic macrolichens. We used non-metric multidimensional scaling to extract gradients in lichen community composition from surveys at 115 forested sites. A strong ammonia deposition gradient was detected, as evidenced by a high linear correlation with an index of ammonia indicator species conventionally known as “nitrophytes” (r = 0.93). This gradient, however, was confounded by elevation (r = ?0.54). We evaluated three statistical techniques for controlling the influence of elevation on nitrophytes: simple linear regression, nonlinear regression, and nonparametric regression. We used the unstandardized residuals from nonlinear regression to estimate relative ammonia deposition at each plot, primarily because this model had the best fit (r ² = 0.33), desirable asymptotic properties, and it is easy to apply to new data. Other possible sources of noise in the nitrophyte-ammonia relationship, such as substrate pH and acidic deposition, are discussed. Lichen communities indicated relatively high deposition to forests of the southern Sierra Nevada, the Modoc Plateau, as well as in stands near urban areas. Evidence of elevated ammonia was also detected for popular recreation areas such as Sequoia and Yosemite National Parks. Lichen communities from forests in the Tahoe basin, northern Sierra Nevada, southern Cascades, and eastern Klamath Range appeared considerably less impacted. This model will be used for continual assessment of eutrophication risks to forest health in the region.     

The effect on pathogenesis of Newcastle disease virus LaSota strain from a mutation of the fusion cleavage site to a virulent sequence

The principal molecular determinant of virulence of Newcastle disease virus (NDV) is the amino acid sequence at the fusion cleavage activation site. To extend the understanding of the role of the fusion cleavage activation site in NDV virulence, the pathogenesis in chickens of a lentogenic LaSota isolate and two infectious clones, NDFL and NDFLtag, were compared. NDFL is an infectious clone of a lentogenic NDV strain (LaSota E13-1), and NDFLtag is the infectious clone with the fusion cleavage site sequence mutated to the virulent motif. NDFL and NDFLtag were described by Peeters et al. The viruses were inoculated intraconjunctivally into groups of 4-wk-old white leghorn chickens and compared in a pathogenesis study for determination of disease causation (clinical signs of disease, gross lesions, histology, virus isolation, and serology) and viral distribution (presence of viral nucleoprotein and mRNA was detected by immunohistochemistry and in situ hybridization, respectively). The modification of the fusion cleavage activation site to the virulent motif in the infectious clone only slightly increased disease severity and viral distribution in the pathogenesis assessment, even though dramatically increased pathogenicity of NDFLtag was confirmed by standard pathogenicity index tests. The result, that the mutated fusion cleavage site of NDV-NDFLtag had only a small influence on pathogenesis in chickens compared to either E13-1 or NDFL, suggests that the pathogenic effects of NDV are not dependent on the fusion cleavage site alone.     

Cloning and characterization of two glutathione S-transferases from pyrethroid-resistant Culex pipiens

BACKGROUND: The Marin strain of Culex pipiens Say is a pyrethroid‐resistant population that was collected in Marin County, California, in 2001 and subsequently maintained in the laboratory under regular permethrin exposure. RESULTS: In this study, two cDNAs, CpGSTd1 and CpGSTd2, encoding glutathione S‐transferase (GST) were cloned from Cx. pipiens Marin. Phylogenetic analysis of the deduced amino acid sequences, CpGSTD1 and CpGSTD2, of these genes indicated that they belong to the Delta class of insect GSTs. The nucleotide and deduced amino acid sequences of CpGSTd1 and CpGSTd2 were 59 and 48% identical respectively. CpGSTD1 and CpGSTD2 were expressed in Escherichia coli and purified by affinity chromatography. The recombinant GSTs exhibited unique selectivity towards the general GST substrates 1‐chloro‐2,4‐dinitrobenzene (CDNB) and 1,2‐dichloro‐4‐nitrobenzene (DCNB), and also differed in their sensitivity to known inhibitors of GSTs. CpGSTD1 exhibited peroxidase activity with cumene hydroperoxide, while CpGSTD2 appeared to lack this activity. CpGSTD1 was able to metabolize 1,1,1‐trichloro‐2,2‐bis(4‐chlorophenyl)ethane (DDT), while DDT metabolism by CpGSTD2 was not detectable. CpGSTD1 and CpGSTD2 showed no detectable metabolism of permethrin. Gene expression of CpGSTd1 and CpGSTd2 in Marin mosquitoes was elevated about twofold in comparison with that found in a pyrethroid‐sensitive mosquito strain. CONCLUSION: The results indicate that CpGSTD1 and CpGSTD2 have unique biochemical characteristics, but they do not appear to play major roles in permethrin resistance in Marin mosquitoes. Copyright © 2012 Society of Chemical Industry     

Early harvest and ensilage of forage sorghum infected with ergot (Claviceps africana) reduces the risk of livestock poisoning

Sorghum ergot produces dihydroergosine (DHES) and related alkaloids, which cause hyperthermia in cattle. Proportions of infected panicles (grain heads), leaves and stems were determined in two forage sorghum crops extensively infected 2 to 4 weeks prior to sampling and the panicles were assayed for DHES. Composite samples from each crop, plus a third grain variety crop, were coarsely chopped and half of each sealed in plastic buckets for 6 weeks to simulate ensilation. The worst-infected panicles contained up to 55 mg DHES/kg, but dilution reduced average concentrations of DHES in crops to approximately 1 mg/kg, a relatively safe level for cattle. Ensilation significantly (P = 0.043) reduced mean DHES concentrations from 0.85 to 0.46 mg/kg.