Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

Development of biomarkers inAporrectodea caliginosa andLumbricus rubellus for detecting exposure to benzo[a]pyrene

Objectives and Methods. The sensitivity of two biomarkers, the neutral red retention assay (NRRA) and cytochrome P450 were evaluated in the earthwormAporrectodea caliginosa for their potential to detect exposure to benzo[a]pyrene (BaP). Cytochrome P450 was also evaluated in the earthwormLumbricus rubellus, and measured using the substrate ethoxycoumarin. Optimal assay conditions (pH, and temperature) were determined, followed by exposure of earthworms to 20 mg/kg BaP (a typical concentration at contaminated sites in New Zealand). Ethoxycoumarin-O-dealkylase (ECOD) activity was measured after 7, 14, and 21 days exposure. The NRRA was evaluated in earthworms exposed to 0.2, 20, and 100 mg BaP/kg, and biomarker responses were compared with effects on body weight. Results and Discussion Benzo[a]pyrene failed to induce ECOD activity in either earthworm species, and therefore it is not useful as a biomarker of BaP exposure and was not evaluated further. In all cases, the NRRA was significantly affected in the absence of any effects on earthworm body weight, indicating that this assay can detect exposure to BaP at a range of concentrations comparable to those found at contaminated sites. The NRRA should be linked to reproductive endpoints, then it can be used as an early warning indicator of adverse effects. Establishing biomarker stability under environmental conditions is an important step in biomarker development. Therefore, the effects of soil type (sandy soil, silt loam, and a clay soil), moisture content (15–30%), and temperature (5–20°) on the NRRA were determined. In all cases, there was no effect on the NRRA, indicating that this assay is very stable under varying environmental conditions. Conclusion and Outlook In conclusion, cytochrome P450 activity does not appear to be a useful indicator of PAH exposure in eitherA. caliginosa or L.rubellus, and due to the inherently low activity, it is not suitable as a routine biomarker for detecting environmental contamination by these compounds. In contrast, the NRRA in the earthworm A.caliginosa is a promising indicator of PAH exposure at the concentrations likely to be found in contaminated sites in New Zealand, and therefore has potential for evaluating these contaminated sites. If the NRRA can be linked to ecologically important life-cycle endpoints, such as reproduction, then it could be used as an early warning indicator of adverse effects at contaminated sites, i.e., by measuring biomarker responses in earthworms from a ‘contaminated area’ and comparing these with earthworms from a matched control area.     

Apium graveolens PI 181714 is a source of resistance to Fusarium oxysporum f. sp. apii race 4 in celery (A. graveolens var. dulce)

Celery has little genetic diversity and is highly susceptible to the new fungal pathogen Fusarium oxysporum f. sp. apii (Foa) race 4. After screening an Apium graveolens germplasm collection for resistance to Foa race 4, we crossed celery cv. 'Challenger', which is Foa race 2-resistant but Foa race 4-susceptible and A. graveolens PI 181714, which is Foa races 2- and 4-resistant but non-celery type. After selfing F₁s, we screened the F₁S₁ for race 4-resistance and celery-type and then selfed selected F₁S₁. Greenhouse and field trials indicate that three selected F₁S₂ families (76–8-4, 76–8-27 and 76–8-36) are suitable as germplasm for celery breeders for resistance to Foa race 4. A F1S3 76–8–36-124 is either fixed or nearly so for resistance to Foa races 4 and 2. Furthermore, quantitative PCR indicates that PI 181714 is resistant, rather than tolerant, to Foa races 4 and 2, and that this resistance has been introgressed into F1S3 76–8–36-124.     

Using EM and VERIS technology to assess land suitability for orchard and vineyard development

Orchard and vineyard producers conduct preplant site evaluations to help prevent planting permanent tree and vine crops on lands where the crop will not perform to its highest potential or attain its full life expectancy. Physical soil characteristics within specific soil profiles and spatially throughout an orchard influence decisions on land preparation, irrigation system selection, horticultural choices, and nutrient management. Producers depend on soil surveys to help them understand the soil characteristics of the land and may be interested in technology that provides additional information. Electromagnetic induction (EM38) and four-probe soil resistance sensors (VERIS) are being used in combination with global positioning systems to map spatial variability of soils using apparent soil electrical conductivity (ECa). The hypothesis evaluated in this study is whether rapid, in situ, and relatively low-cost methods of measuring ECa (EM38 and VERIS) can effectively identify and map physical soil variability in non-saline soils. The supposition is that in non-saline soils, ECa levels will relate well to soil texture and water-holding capacity and can be used to map physical soil variability. In turn, the information can be used to guide decisions on preplant tillage, irrigation system design, water and nutritional management, and other horticultural considerations. Two sites in the Sacramento Valley were mapped each with EM38 and VERIS methods. Site-specific management zones were identified by each provider on ECa maps for each site, and then soil samples were collected by University of California researchers to verify these zones. Results showed that on non-saline soils, ECa measured with both EM38 and VERIS correlate with physical soil properties such as gravel, sand, silt, and clay content but the relationship between conductivity and these physical soil properties varied from moderately strong to weak. The strength of the correlation may be affected by several factors including how dominant soil texture is on conductivity relative to other soil properties and on methods of equipment operation, data analysis and interpretation. Overall, the commercial providers of ECa surveys in this study delivered reasonable levels of accuracy that were consistent with results reported in previous studies. At one site, an ECa map developed with VERIS provided more detail on physical soil variability to supplement published soil surveys and aided in the planning and development of a walnut orchard. At a second site, almond yield appeared to correlate well with distinctly different soil zones identified with EM38 mapping.     

Field scale spatiotemporal analysis of surface soil moisture for evaluating point-scale in situ networks

Soil moisture is an intrinsic state variable that varies considerably in space and time. From a hydrologic viewpoint, soil moisture controls runoff, infiltration, storage and drainage. Soil moisture determines the partitioning of the incoming radiation between latent and sensible heat fluxes. Although soil moisture may be highly variable in space and time, if measurements of soil moisture at the field or small watershed scale are repeatedly observed, certain locations can often be identified as being temporally stable and representative of the an area average. This study is aimed at determining the adequacy of long term point-scale surface soil moisture measurements in representing local field scale averages which may ultimately serve as in situ locations for the calibration and validation of remotely sensed soil moisture. Experimental data were obtained by frequency-domain reflectometry (FDR) sensors permanently installed in two agricultural fields, AS1 and AS2 (2.23 and 2.71 ha, respectively) at a depth of 5 cm. Twenty additional FDR sensors, spaced 35 m apart, were installed horizontally at a depth of 5 cm in each field with automated data collection being transmitted every 30 min from July 15 through September 20, 2009. Additionally, meteorological data were obtained from existing weather stations in each field. The FDR sensors revealed persistent patterns in surface soil moisture within each field and identified sites that were temporally stable. The locations that were optimal for estimating the area-average field water contents were different from the permanent sensor locations in both fields. Permanent sensor data showed approximately 4 and 10% mean relative differences for fields AS1 and AS2, respectively, with relatively large standard deviations. Thus, minimum offset values could be applied to the temporally stable field sites to obtain representative field average values of surface soil moisture. However, use of permanent sensor data for offset estimates gave poor results. These findings are of relevance for applications of geospatial surface soil moisture data assimilation in hydrologic modeling when only point-scale observations are available, as well as, remotely sensed surface soil moisture calibration and validation studies.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.     

Intestinal microbiota contribute to the endogenous formation of thiouracil in livestock

In recent years, the frequent detection of the banned substance thiouracil (TU) in livestock urine has been related to its endogenous formation following consumption of glucosinolate-rich Brassicaceae crops. Besides, TU residues have been recovered in these crops upon plant-derived myrosinase hydrolysis. Through in vitro bovine and porcine static digestive simulations, the influence of gastrointestinal digestion of Brassicaceae-derived matrixes on TU formation was assessed. Following derivatization and LC-MS(2) analysis, TU was detected in colonic suspensions with traditional rapeseed, coarse colza "00" meal, cauliflower, and broccoli ranging from 3.47 to 30.96 μg kg(-1) (bovine) and from 3.55 to 26.34 μg kg(-1) (porcine). In stomach and small intestinal fluids, TU remained unfound, whereas upon rumen simulation TU was detected for coarse colza "00" meal (4.43 μg kg(-1)) and grounded traditional rapeseed (0.35 μg kg(-1)). The origin of this detection was investigated through filter-sterilizing and autoclaving the fecal inoculum causing a significant decrease in TU concentration, thereby reinforcing the possibility of an active bacterial involvement, which however was characterized with a high interanimal variation. In conclusion, these results support the previously proven endogenous origin of TU and acknowledge the active role of the gastrointestinal bacteria in TU formation, through production of an extracellular component.     

Feed or food responsible for the presence of low-level thiouracil in urine of livestock and humans?

In recent years, questions have been raised on the possible semi-endogenous status of the alleged xenobiotic thyreostatic drug thiouracil; thiouracil has been detected in the urine of various animals (livestock and domesticated) at concentrations between 1 and 10 μg L(-1) and also in human urine. Although several studies suggest Brassicaceae-derived feed as potential origin, no traces of thiouracil have been detected in feed so far. Therefore, the aim of this study was to elucidate the origin of thiouracil in the urine of livestock and humans. To this purpose various Brassicaceae feed and food sources (e.g., rapeseed, rapeseed coarse meal, cabbage, cauliflower, broccoli) were investigated for the presence of thiouracil. In addition, the impact of the Brassicaceae-related β-thioglucosidase enzyme was evaluated. This myrosinase enzyme appeared to be crucial, because without its catalyzed hydrolysis no thiouracil could be detected in the various Brassicaceae-derived samples. Therefore, a sample pretreatment with incorporated enzymatic hydrolysis was developed after ensuring the quality performance of the extracted myrosinase mixture with a single-point glucose assay. Upon enzymatic hydrolysis and LC-MS(2) analysis, thiouracil was successfully detected in samples of traditional rapeseed, rapeseed-'00' variety coarse meal (values of erucic acid <2% and glucosinolates <25 μmol g(-1)), and rapeseed cake at 1.5, 1.6, and 0.4 μg kg(-1), respectively. As for the food samples, broccoli and cauliflower displayed thiouracil concentrations of 6.0 and <1.0 μg kg(-1), respectively. To the best of the authors' knowledge this study is the first to report the presence of naturally occurring thiouracil in feed and food samples. Future research should investigate the pathway of thiouracil formation and identify its possible precursors.