In vitro screening of technical lignins to determine their potential as hay preservatives.

Our objectives were to evaluate technical lignins for their antifungal properties against 3 molds and 1 yeast causing hay spoilage, and their ability to preserve ground high-moisture alfalfa hay nutritive value in vitro. In experiment 1, 8 technical lignins and propionic acid (PRP; positive control) were tested at a dose of 40 mg/mL. The experiment had a randomized complete block design (RCBD, 4 runs) and a factorial arrangement of 3 molds × 10 additives (ADV). The effects of the ADV on yeast were evaluated separately with a RCBD. Sodium lignosulfonate (NaL) and PRP were the only treatments with 100 ± 2.8% inhibition of fungi. In experiment 2, the minimum inhibitory concentration (MIC) for selected lignins and PRP were determined. At pH 4, NaL had the lowest MIC across the molds (20-33.3 mg/mL) and magnesium lignosulfonate (MgL) for the yeast (26.7) among the lignins. However, PRP had MIC values that were several-fold lower across all fungi (1.25-3.33). In experiment 3, a RCBD (5 blocks) with a 3 (ADV; NaL, MgL, and PRP) × 4 (doses: 0, 0.5, 1, and 3% wt/wt fresh basis) factorial arrangement of treatments was used to evaluate the preservative effects of ADV in ground high-moisture alfalfa hay inoculated with a mixture of the fungi previously tested and incubated under aerobic conditions in vitro. After 15 d, relative to untreated hay (14.9), dry matter (DM) losses were lessened by doses as low as 1% for NaL (3.39) and 0.5% for PRP (0.81 ± 0.77%). The mold count was reduced in both NaL at 3% (3.92) and PRP as low as 0.5% (3.94) relative to untreated hay (7.76 ± 0.55 log cfu/fresh g). Consequently, sugars were best preserved by NaL at 3% (10.1) and PRP as low as 0.5% (10.5) versus untreated (7.99 ± 0.283% DM), while keeping neutral detergent fiber values lower in NaL (45.9) and PRP-treated (45.1) hays at the same doses, respectively, relative to untreated (49.7 ± 0.66% DM). Hay DM digestibility was increased by doses as low as 3% for NaL (67.5), 1% MgL (67.0), and 0.5% PRP (68.5) versus untreated hay (61.8 ± 0.77%). The lowest doses increasing neutral detergent fiber digestibility relative to untreated hay (23.3) were 0.5% for MgL and PRP (30.5 and 30.1, respectively) and 1% for NaL (30.7 ± 1.09% DM). Across technical lignins, NaL showed the most promise as a potential hay preservative. However, its effects were limited compared with PRP at equivalent doses. Despite not having an effect on preservation, MgL improved DM digestibility by stimulating neutral detergent fiber digestibility. This study warrants further development of NaL under field conditions.

Poster - Thur Eve - 76: A quality control to achieve planning consistency in arc radiotherapy of the prostate.

PURPOSE: To report a quality control program in prostate radiation therapy at our center that includes semi-automated planning process to generate high quality plans and in-house software to track plan quality in the subsequent clinical application. MATERIAL AND METHODS: Arc planning in Eclipse v10.0 was preformed for both intact prostate and post-prostatectomy treatments. The planning focuses on DVH requirements and dose distributions being able to tolerate daily setup variations. A modified structure set is used to standardize the optimization, including short rectum and bladder in the fields to effectively tighten dose to target and a rectum expansion with 1cm cropped from PTV to block dose and shape posterior isodose lines. Structure, plan and optimization templates are used to streamline plan generation. DVH files are exported from Eclipse to a quality tracking software with GUI written in Matlab that can report the dose-volume data either for an individual patient or over a patient population. RESULTS: For 100 intact prostate patients treated with 78Gy, rectal D50, D25, D15 and D5 are 30.1±6.2Gy, 50.6±7.9Gy, 65.9±6.0Gy and 76.6±1.4Gy respectively, well below the limits 50Gy, 65Gy, 75Gy and 78Gy respectively. For prostate bed with prescription of 66Gy, rectal D50 is 35.9±6.9Gy. In both sites, PTV is covered by 95% prescription and the hotspots are less than 5%. CONCLUSION: The semi-automated planning method can efficiently create high quality plans while the tracking software can monitor the feedback from clinical application. It is a comprehensive and robust quality control program in radiation therapy.

Poster - Thur Eve - 54: A software solution for ongoing DVH quality assurance in radiation therapy.

PURPOSE: A program has been developed in MATLAB for use in quality assurance of treatment planning of radiation therapy. It analyzes patient DVH files and compiles dose volume data for review, trending, comparison and analysis. MATERIAL AND METHODS: Patient DVH files are exported from the Eclipse treatment planning system and saved according to treatment sites and date. Currently analysis is available for 4 treatment sites; Prostate, Prostate Bed, Lung, and Upper GI, with two functions for data report and analysis: patient-specific and organ-specific. The patient-specific function loads one patient DVH file and reports the user-specified dose volume data of organs and targets. These data can be compiled to an external file for a third party analysis. The organ-specific function extracts a requested dose volume of an organ from the DVH files of a patient group and reports the statistics over this population. A graphical user interface is utilized to select clinical sites, function and structures, and input user's requests. RESULTS: We have implemented this program in planning quality assurance at our center. The program has tracked the dosimetric improvement in GU sites after VMAT was implemented clinically. It has generated dose volume statistics for different groups of patients associated with technique or time range. CONCLUSION: This program allows reporting and statistical analysis of DVH files. It is an efficient tool for the planning quality control in radiation therapy.

Leaf and Flower Blight Caused by Monilinia vaccinii-corymbosi on Lowbush Blueberry: Effects on Yield and Relationship to Bud Phenology.

ABSTRACT Naturally established lowbush blueberry clones in four fields were evaluated for the incidence of leaf and flower blight, proportion of mummy berries, and yield reductions caused by Monilinia vacciniicorymbosi. The relationship between the phenology of flower and leaf bud development and susceptibility also was examined. Three fields were examined over one crop year and one field was studied in two subsequent crop years. The incidence of stems with blight was correlated to incidence of leaf blight in all fields and to incidence of flower blight in one field. Incidence of leaf and flower blight and the proportion of mummy berries produced were not correlated. Lowbush blueberry clones with higher incidence levels of leaf blight had reduced fruit set and lower berry weights. For healthy stems, leaf-to-fruit ratios had no effect on berry weight in most fields. In contrast, blighted stems with higher leaf-to-fruit ratios had higher berry weights in three fields. Stems with slowerdeveloping leaf and flower buds had less leaf and flower blight, respectively, than stems with faster bud development. Some blueberry clones may avoid infection by delaying production of susceptible tissue until after ascospore release by M. vaccinii-corymbosi.

Novel procedure for identification of compounds inhibitory to transcription of genes involved in mycotoxin biosynthesis.

A novel assay is described for the identification and isolation of compounds that inhibit the transcription of genes involved in mycotoxin biosynthesis. The thin-layer chromatography-based assay was used to screen plant extracts for compounds that would inhibit the expression of the beta-glucuronidase reporter gene under the control of an aflatoxin biosynthesis gene promoter in Aspergillus parasiticus. The assay was used to track purification of an inhibitory compound, cp2, from extracts of black pepper (Piper nigrum). Cp2 did not inhibit mycelial growth or the expression of the beta-tubulin gene but did inhibit aflatoxin biosynthesis at the transcriptional level. Applications of cp2 to the control of mycotoxins are discussed.

Presence of peptide synthetase gene transcripts and accumulation of ergopeptines in Claviceps purpurea and Neotyphodium coenophialum.

The production of toxic ergopeptine alkaloids by the fungi Claviceps purpurea and Neotyphodium coenophialum involves the activity of one or more nonribosomal peptide synthetases. Claviceps purpurea and N. coenophialum each have several different peptide synthetase genes, fragments of which have been cloned previously. An additional Claviceps purpurea peptide synthetase gene was cloned by hydridization with one of the N. coenophialum peptide synthetase gene fragments. We detected the presence of mRNA from the peptide synthetase genes in cultures of different ages grown under conditions favorable or unfavorable for ergopeptine production. All four peptide synthetase genes from Claviceps purpurea were transcribed under at least some of the experimental conditions. Transcripts from three of the four genes were detected under conditions consistent with their potential involvement in ergopeptine biosynthesis. All three peptide synthetase genes previously identified in N. coenophialum were transcribed during symbiotic growth of this fungus with tall fescue, as well as ergopeptine-producing cultures. The data show that all of the peptide synthetase genes are transcribed, that one of the peptide synthetase genes is dissociated from ergopeptine biosynthesis, and, as a result, prioritize the remaining genes for functional analyses by transformation-mediated gene disruption.

Transposon-like sequences at the TOX2 locus of the plant-pathogenic fungus Cochliobolus carbonum.

The ascomycete fungus Cochliobolus carbonum race 1 is pathogenic on certain genotypes of maize due to the production of HC-toxin, a host-specific cyclic peptide. HC-toxin production is controlled, at least in part, by a duplicated 22-kb region of DNA that is found only in toxin-producing isolates of the fungus. This 22-kb region of DNA is flanked by a repetitive element. We have sequenced the element and found an interrupted reading frame that would encode a product similar to transposases from the fungal transposons Fot1 of Fusarium oxysporum and Pot2 of Magnaporthe grisea. The individual element cloned from C. carbonum is likely to function neither in cis nor trans, as it had a nonsense mutation in frame and several substitutions in its terminal inverted repeats. However, similar elements in the C. carbonum genome may be active, as the putative transposase-encoding region hybridized to mRNA of the size predicted by the reading frame. The element was found in varying copy number in the genomes of all Cochliobolus spp. examined, giving a distinct fingerprint in each species and race tested. The sequence similarity of the C. carbonum repetitive element to other fungal transposons, along with its presence in multiple copies per genome, strongly suggest that the C. carbonum repetitive element is a member of the Fot1 family of fungal transposons.

Rapid identification of genetic variation and pathotype of Leptosphaeria maculans by random amplified polymorphic DNA assay.

Canadian isolates of Leptosphaeria maculans, the causal agent of blackleg of crucifers, were examined for genetic relatedness by the random amplified polymorphic DNA assay. DNA polymorphisms amplified with random decamer primers were used to distinguish three groups of isolates. Group 1 contained all isolates of the virulent pathotype, group 2 contained isolates of the avirulent pathotype from western Canada, and group 3 contained avirulent pathotype isolates from Ontario. These results agreed with other reports which showed many genetic differences between pathotypes and were consistent with the hypothesis that the virulent pathotype was recently introduced into Canada and has diverged relatively little. In contrast, the avirulent pathotype has probably been present in Canada for a longer time and has diverged with geographic isolation. In addition to establishing genetic relationships, DNA fingerprints generated by the random amplified polymorphic DNA assay have potential applications in pathotype identification and blackleg disease management.