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1.
Phytopathology ; 113(12): 2187-2196, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37287124

ABSTRACT

Pantoea vagans C9-1 (C9-1) is a biological control bacterium that is applied to apple and pear trees during bloom for suppression of fire blight, caused by Erwinia amylovora. Strain C9-1 has three megaplasmids: pPag1, pPag2, and pPag3. Prior bioinformatic studies predicted these megaplasmids have a role in environmental fitness and/or biocontrol efficacy. Plasmid pPag3 is part of the large Pantoea plasmid (LPP-1) group that is present in all Pantoea spp. and has been hypothesized to contribute to environmental colonization and persistence, while pPag2 is less common. We assessed fitness of C9-1 derivatives cured of pPag2 and/or pPag3 on pear and apple flowers and fruit in experimental orchards. We also assessed the ability of a C9-1 derivative lacking pPag3 to reduce populations of E. amylovora on flowers and disease incidence. Previously, we determined that tolerance to stresses imposed in vitro was compromised in derivatives of C9-1 lacking pPag2 and/or pPag3; however, in this study, the loss of pPag2 and/or pPag3 did not consistently reduce the fitness of C9-1 on flowers in orchards. Over the summer, pPag3 contributed to survival of C9-1 on developing apple and pear fruit in two of five trials, whereas loss of pPag2 did not significantly affect survival of C9-1. We also found that loss of pPag3 did not affect C9-1's ability to reduce E. amylovora populations or fire blight incidence on apple flowers. Our findings partially support prior hypotheses that LPP-1 in Pantoea species contributes to persistence on plant surfaces but questions whether LPP-1 facilitates host colonization.


Subject(s)
Erwinia amylovora , Malus , Pantoea , Pyrus , Malus/microbiology , Fruit , Pantoea/genetics , Pyrus/microbiology , Plant Diseases/prevention & control , Plant Diseases/microbiology , Plasmids , Erwinia amylovora/genetics , Flowers/microbiology
2.
Phytopathology ; 113(12): 2205-2214, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37530490

ABSTRACT

The stability of the fire blight control material, oxytetracycline, in water is strongly affected by pH, increasing with increasing acidity. From 2017 to 2021, pear and apple orchard trials were conducted to evaluate if acidic amendments to oxytetracycline sprays improve fire blight control. Compared with the water-treated control, infection suppression after two bloom applications of an acidified commercial oxytetracycline formulation averaged 85.9 ± 0.4% compared with 72.2 ± 1.7% without an acidifier, but individual trials frequently had insufficient statistical power to separate among acidified and non-acidified antibiotic treatments. Across trials, a significant linear relationship was observed for regression of relative infection suppression from oxytetracycline (hydrochloride formulation) on spray tank pH. Similar relationships were observed for oxytetracycline (calcium complex formulation) and kasugamycin (P values were 0.055 and 0.069, respectively). Also based on regression, acidified oxytetracycline and kasugamycin suppressed epiphytic populations of Erwinia amylovora on flowers to a greater degree than the antibiotic only. As spray suspensions, commercial oxytetracycline formulations at label rate and amended with citric acid (1.2 g/liter) in well water had pH values near 3.4, but after spraying, the pH of flowers washed in deionized water (1 ml/flower) measured in a range of 5.2 to 5.5 compared with a pH range of 5.8 to 6.0 after a treatment of oxytetracycline only. In pear fruit finish trials, sprays acidified with citric acid-based materials had negligible effects on fruit russeting. Based on a serological assay, the detectable residual of oxytetracycline on apple foliage was increased by co-application with citric acid compared with a non-acidified control.


Subject(s)
Erwinia amylovora , Malus , Oxytetracycline , Pyrus , Oxytetracycline/pharmacology , Plant Diseases/prevention & control , Anti-Bacterial Agents/pharmacology , Citric Acid , Water
3.
Phytopathology ; 113(12): 2197-2204, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37344783

ABSTRACT

Fire blight, caused by Erwinia amylovora, is an economically important disease in apples and pears worldwide. This pathogen relies on the type III secretion system (T3SS) to cause disease. Compounds that inhibit the function of the T3SS (T3SS inhibitors) have emerged as alternative strategies for bacterial plant disease management, as they block bacterial virulence without affecting growth, unlike traditional antibiotics. In this study, we investigated the mode of action of a T3SS inhibitor named TS108, a plant phenolic acid derivative, in E. amylovora. We showed that adding TS108 to an in vitro culture of E. amylovora repressed the expression of several T3SS regulon genes, including the master regulator gene hrpL. Further studies demonstrated that TS108 negatively regulates CsrB, a global regulatory small RNA, at the posttranscriptional level, resulting in a repression of hrpS, which encodes a key activator of hrpL. Additionally, TS108 has no impact on the expression of T3SS in Dickeya dadantii or Pseudomonas aeruginosa, suggesting that its inhibition of the E. amylovora T3SS is likely species specific. To better evaluate the performance of T3SS inhibitors in fire blight management, we conducted five independent field experiments in four states (Michigan, New York, Oregon, and Connecticut) from 2015 to 2022 and observed reductions in blossom blight incidence as high as 96.7% compared with untreated trees. In summary, the T3SS inhibitors exhibited good efficacy against fire blight.


Subject(s)
Erwinia amylovora , Malus , Type III Secretion Systems/genetics , Type III Secretion Systems/metabolism , Erwinia amylovora/genetics , Erwinia amylovora/metabolism , Plant Diseases/prevention & control , Plant Diseases/microbiology , Anti-Bacterial Agents/pharmacology , Malus/microbiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism
4.
Phytopathology ; 113(7): 1192-1201, 2023 Jul.
Article in English | MEDLINE | ID: mdl-36794987

ABSTRACT

Fire blight, caused by Erwinia amylovora, is a devastating disease of apple. Blossom Protect, a product that contains Aureobasidium pullulans as the active ingredient, is one of the most effective biological controls of fire blight. It has been postulated that the mode of action of A. pullulans is to compete against and antagonize epiphytic growth of E. amylovora on flowers, but recent studies have found that flowers treated with Blossom Protect harbored similar to or only slightly reduced E. amylovora populations compared with nontreated flowers. In this study, we tested the hypothesis that A. pullulans-mediated biocontrol of fire blight is the result of induced host resistance. We found that PR genes in the systemic acquired resistance pathway, but not genes in the induced systemic resistance pathway, were induced in hypanthial tissue of apple flowers after the Blossom Protect treatment. Additionally, the induction of PR gene expression was coupled with an increase of plant-derived salicylic acid in this tissue. After inoculation with E. amylovora, PR gene expression was suppressed in nontreated flowers, but in flowers pretreated with Blossom Protect, the heightened PR expression offset the immune repression caused by E. amylovora, and prevented infection. Temporal and spatial analysis of PR gene induction showed that induction of PR genes occurred 2 days after the Blossom Protect treatment, and required direct flower-yeast contact. Finally, we observed deterioration of the epidermal layer of the hypanthium in some of the Blossom Protect-treated flowers, suggesting that PR gene induction in flowers may be a result of pathogenesis by A. pullulans.


Subject(s)
Malus , Malus/genetics , Plant Diseases/genetics , Flowers , Gene Expression
5.
Plant Dis ; 106(6): 1660-1668, 2022 Jun.
Article in English | MEDLINE | ID: mdl-34854760

ABSTRACT

Winter squash (Cucurbita maxima cultivar Golden Delicious) produced in Oregon's Willamette Valley for edible seed production has experienced significant yield losses because of a soilborne disease. The symptoms associated with this disease problem include root rot, crown rot, and vascular discoloration in the stems, leading to a severe late season wilt and plant collapse. Through field surveys, Fusarium oxysporum, F. solani, F. culmorum-like fungi, Plectosphaerella cucumerina, and Setophoma terrestris were identified to be associated with diseased tissues, and each produced symptoms of root rot, crown rot, or stem discoloration in preliminary pathogenicity trials. In this study, 219 isolates of these species were characterized by molecular identity analyses using BLAST of the internal transcribed spacer and translation elongation factor 1 alpha genomic regions and by pathogenicity testing in outdoor, large-container trials. Molecular identity analyses confirmed the identity of isolates at 99 to 100% similarity to reference isolates in the database. In pathogenicity experiments, F. solani produced the most severe symptoms, followed by F. culmorum-like fungi, F. oxysporum, P. cucumerina, and S. terrestris. Some treatments of mixed-species inoculum produced symptom severity greater than what was expected from individual species. In particular, the mixture of F. culmorum-like fungi, F. oxysporum, and P. cucumerina and the mixture of F. culmorum-like fungi, F. solani, and S. terrestris had symptom ratings as high as that of F. solani by itself. Results indicate that this soilborne disease is caused primarily by Fusarium solani, but interactions between the complex of F. solani, F. culmorum-like fungi, F. oxysporum, and P. cucumerina can exacerbate disease severity.


Subject(s)
Cucurbita , Plant Diseases , Plant Diseases/microbiology , Virulence
6.
Plant Dis ; 106(2): 623-633, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34633232

ABSTRACT

Fire blight-susceptible, certified organic pome fruit is produced on 9,000 ha in the Pacific Northwest region of the United States with acreage continuing to expand despite a 2014 prohibition on antibiotics as allowable materials for infection suppression. Nonantibiotic practices for fire blight pathogen suppression mirror conventional management, but the full-bloom-to-petal-fall period when antibiotics are typically sprayed for fire blight control continues to receive research scrutiny owing to drawbacks and weaknesses of alternative materials. As solitary treatments, effective nonantibiotic materials (e.g., a yeast biocontrol, soluble coppers, and potassium aluminum sulfate) raise the risk of a crop-value-reducing, phytotoxic response termed "fruit russeting." Conversely, materials with less russeting risk (e.g., Bacillus-based biorationals) are less effective for fire blight control. Spray programs using a sequence of materials applied from midbloom to petal fall have the potential to provide high levels of protection with reduced russeting risk. In orchard trials, the effects of nonantibiotic spray programs on the epiphytic population size of Erwinia amylovora in flowers, yeast biocontrol population size, floral pH, infection suppression, and fruit russeting revealed strategies for sequencing sprays of nonantibiotic materials. The yeast biocontrol, Blossom Protect (Aureobasidium pullulans), sprayed at 70% bloom, was an important contributor to fire blight pathogen suppression as was the soluble copper material, Previsto, when applied at full bloom. Choice of material for the petal-fall spray timing was important to fruit russeting risk but apparently less important to overall infection incidence. Consequently, treatment programs of Blossom Protect at 70% bloom, a soluble copper at full bloom, and a Bacillus-based biorational at petal fall, best balance the quality of infection suppression with the risk of fruit russeting.


Subject(s)
Erwinia amylovora , Malus , Flowers , Fruit , Plant Diseases/prevention & control
7.
Plant Dis ; 101(7): 1263-1268, 2017 Jul.
Article in English | MEDLINE | ID: mdl-30682961

ABSTRACT

Induction of systemic acquired resistance as a therapeutic aid to restoration of tree health was evaluated in 3- to 14-year-old pear and apple trees diseased with fire blight. Acibenzolar-S-methyl (ASM) was applied to diseased trees in late spring near the time of removal of primary fire blight cankers, which had originated from floral infection. Suspensions of ASM (7.5 to 22.5 g of active ingredient per liter plus silicone surfactant) were painted onto a 30- to 45-cm length of branch tissue immediately below primary pruning cuts or sprayed onto an 80- to 100-cm length of central trunk. In some experiments, a second ASM treatment was made in late June to early July in conjunction with secondary pruning of redeveloped cankers. After pruning primary cankers, effects of ASM were measured by assessing weight and length of secondary cankers that were the result of fire blight redevelopment. Over 5 years of field experiments, trees that received an ASM treatments yielded 62% less diseased wood at the time of secondary and tertiary canker removal compared with non-ASM-treated trees. Moreover, tree mortality and proportion of pruning cuts where fire blight redeveloped were reduced by ASM. Induction of systemic acquired resistance could prove practical as an aid to pruning therapy in young, fire-blight-susceptible pear and apple trees where, after canker removal, disease symptoms frequently redevelop owing to residual cells of the pathogen distributed within symptomless portions of the tree.

8.
Plant Dis ; 100(6): 1125-1131, 2016 Jun.
Article in English | MEDLINE | ID: mdl-30682288

ABSTRACT

Greenhouse-grown, 1-year-old potted 'Bosc' pear and apple rootstock cultivars 'M.9' and 'M.26' were inoculated with the fire blight pathogen, Erwinia amylovora, and subjected to trunk paint, root drench, or foliar spray treatments with acibenzolar-S-methyl (ASM, 4 to 30 mg a.i./tree) to induce systemic acquired resistance. Each method of ASM treatment suppressed fire blight canker expansion by 22 to 25%. Furthermore, ASM application method and ASM treatment timing (at or ±3 weeks relative to inoculation) interacted significantly (P ≤ 0.02) in each experiment. A root drench was most effective when applied 3 weeks before inoculation (36% suppression) whereas trunk paints and foliar sprays were more effective at inoculation (43 and 34%, suppression, respectively). Sizes of fire blight cankers in potted apple rootstocks M.9 and M.26 (under scions 'Gala' or 'Cameo') inoculated directly with the pathogen were reduced by 82 and 87% after two pretreatments of ASM applied as a trunk paint or root drench, respectively. Expression of pathogenesis-related (PR) genes PR-1 and -2 in apple leaves sampled after an ASM trunk paint were elevated significantly (P ≤ 0.05) relative to control trees for at least 9 weeks after treatment. Results of this study are being used to guide field research on postinfection therapy with ASM in 1- to 10-year-old pear and apple trees where fire blight has proven difficult to manage with therapeutic pruning only.

9.
Plant Dis ; 99(8): 1147-1152, 2015 Aug.
Article in English | MEDLINE | ID: mdl-30695942

ABSTRACT

From 2010 to 2013, the efficacy of copper-based inoculum sanitation as a component of fire blight management programs was evaluated in commercial pear orchards located in northern California. Forty-one 4-ha sections of orchard were split into two equal-sized plots, with the orchardist applying horticultural oil alone to one plot and horticultural oil plus a fixed copper bactericide to the other plot. These treatments were timed to begin just prior to and finish at the "green tip" phenological stage, which occurs about 5 weeks before full bloom. During bloom, flower samples were collected from the plots and subjected to a loop-mediated isothermal DNA amplification (LAMP) assay for specific detection of Erwinia amylovora. Overall, epiphytic populations of E. amylovora on flowers were detected rarely at midbloom (6% of samples) but commonly at petal fall (44% of samples). In three of four seasons, E. amylovora detection in flower samples at a given bloom stage was significantly suppressed in copper-plus-oil-treated plots compared with oil-only plots. All orchards also received antibiotic treatments during the bloom period and, perhaps as a consequence, the development of fire blight was sporadic and not affected significantly by the copper treatment in any season. The pathogen detection data indicate that copper sanitation may add value to a fire blight management program by delaying the increase of epiphytic populations of E. amylovora in flowers to the late stages of the bloom period, at which time the number of susceptible flowers declines rapidly.

10.
Plant Dis ; 97(3): 402-409, 2013 Mar.
Article in English | MEDLINE | ID: mdl-30722367

ABSTRACT

Apple and pear produced organically under the U.S. National Organic Program (NOP) standard can be treated with antibiotics for suppression of fire blight caused by Erwinia amylovora. Recent regulatory actions by the NOP, however, have lessened the likelihood of antibiotic use after the 2014 season. In response, western U.S. organic apple and pear stakeholders identified two immediate-need research objectives related to fire blight control: development of effective non-antibiotic control programs based on combinations of registered biological products; and, in apple, integration of these products with lime sulfur, which is sprayed at early bloom to reduce fruit load. In orchard trials in Oregon, increasing the frequency of treatment with biological products improved suppression of floral infection. In apple, fruit load thinning with 2% lime sulfur plus 2% fish oil (LS+FO) at 30 and 70% bloom significantly (P ≤ 0.05) reduced the proportion of blighted flower clusters in four of five orchard trials. Moreover, lime sulfur significantly (P ≤ 0.05) suppressed epiphytic populations of E. amylovora after their establishment on apple flowers. Over four trials, treatment with Aureobasidium pullulans (Blossom Protect) after LS+FO reduced the incidence of fire blight by an average of 92% compared with water only; this level of control was similar to treatment with streptomycin. In three seasons, a spray of a Pantoea agglomerans product after the 70% bloom treatment of LS+FO established the antagonist on a significantly (P ≤ 0.05) higher proportion of flowers compared with a spray of this bacterium before the thinning treatment. Consequently, in apple, biological treatments for fire blight control are not advised until after lime sulfur treatments for fruit load thinning are completed.

11.
Plant Dis ; 97(12): 1585-1592, 2013 Dec.
Article in English | MEDLINE | ID: mdl-30716831

ABSTRACT

Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the live-dead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 101 CFU for pure cultures of X. hortorum pv. carotae, and 102 to 103 CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R2 = 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilution-plating assay.

12.
Plant Dis ; 95(4): 423-430, 2011 Apr.
Article in English | MEDLINE | ID: mdl-30743332

ABSTRACT

Fire blight of pear and apple is frequently an inoculum-limited disease but weather-based forecasting models commonly assume that the pathogen is omnipresent. To improve fire blight risk assessment during flowering, we developed a rapid pathogen detection protocol that uses loop-mediated isothermal amplification (LAMP) to detect DNA of epiphytic Erwinia amylovora on samples of pear and apple flowers. LAMP detected a single flower colonized epiphytically by E. amylovora in a sample of 100 flower clusters (approximately 600 flowers). Samples of 100 flower clusters from orchards (approximately one sample per hectare) were washed and subjected to LAMP, which was completed in 2 h. In three experimental orchards inoculated with E. amylovora, positive LAMP reactions were attained from nine of nine 100-flower cluster samples; pathogen populations in the floral washes averaged 5.2 × 103 CFU per flower as determined by dilution plating. Samples of pear and apple flowers obtained from 60 commercial orchards located in Oregon, Washington, California, and Utah resulted in detection of E. amylovora by LAMP assay from 34 sites, 20 of which developed fire blight. Of samples at early bloom, 10% were positive for epiphytic E. amylovora compared with 28% at petal fall; pathogen density in washes of positive samples averaged 3.2 × 102 CFU per flower. In another 26 orchards, all floral washes were negative for E. amylovora by LAMP and by dilution plating; a light severity of fire blight was observed in 8 of these orchards. Overall, positive detection of epiphytic E. amylovora in commercial orchards by LAMP-based scouting generally occurred at later stages of bloom after heat (risk) units had begun to accumulate, an indication that weather-based forecasting models may be an adequate measure of fire blight risk for many orchardists. Nonetheless, several orchardists communicated that information from the LAMP-based rapid detection protocol resulted in modification of their fire blight management practices.

13.
Phytopathology ; 99(6): 679-88, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19453226

ABSTRACT

Establishment of suppressive populations of bacterial biological control agents on aerial plant surfaces is a critical phase in biologically based management of floral diseases. Periodically, biocontrol agents encounter inhospitable conditions for growth on plants; consequently, tolerance of environmental stresses may contribute to their fitness. In many gram-negative bacteria, including strains of Pseudomonas spp., the capacity to survive environmental stresses is influenced by the stationary phase sigma factor RpoS. This study focused on the role of RpoS in stress response and epiphytic fitness of Pseudomonas fluorescens A506, a well-studied bacterial biological control agent. We detected a frameshift mutation in the rpoS of A506 and demonstrated that the mutation resulted in a truncated, nonfunctional RpoS. Using site-directed mutagenesis, we deleted a nucleotide from rpoS, which then encoded a full-length, functional RpoS. We compared the stress response and epiphytic fitness of A506 with derivative strains having the functional full-length RpoS or a disrupted, nonfunctional RpoS. RpoS had little effect on stress response of A506 and no consistent influence on epiphytic population size of A506 on pear or apple leaves or flowers. Although the capacity of strain A506 to withstand exposure to environmental stresses was similar to that of other fluorescent pseudomonads, this capacity was largely independent of rpoS.


Subject(s)
Bacterial Proteins/genetics , Flowers/microbiology , Mutation , Plant Diseases/microbiology , Pseudomonas fluorescens/genetics , Sigma Factor/genetics , Amino Acid Sequence , Bacterial Proteins/chemistry , Base Sequence , DNA, Plant/chemistry , DNA, Plant/genetics , Environment , Genetic Complementation Test , Molecular Sequence Data , Mutagenesis, Site-Directed , Plasmids/chemistry , Plasmids/genetics , Pseudomonas fluorescens/pathogenicity , Sigma Factor/chemistry
14.
Phytopathology ; 97(10): 1263-73, 2007 Oct.
Article in English | MEDLINE | ID: mdl-18943684

ABSTRACT

ABSTRACT Phytosanitary concerns about fire blight prohibit export of U.S.-grown pears to some countries without this disease. To examine these concerns, we evaluated the potential for co-occurrence of Erwinia amylovora with mature, symptomless winter pear fruit by inoculation experiments and by survey of commercial orchards. Immature pear and apple fruit were inoculated in orchards with E. amylovora strain 153N as resuspended lyophilized cells or as ooze from diseased tissues. Regardless of inoculum source, population size of Ea153N on fruit declined by an order of magnitude every 3 to 4 days during the first 2 weeks after inoculation; at 56 days after inoculation, Ea153N was not detected, except on 1 of 450 fruit with 4 colony forming units (CFU). After inoculation of flowers, calyx-end survival of Ea153N on pear and apple fruit declined from high populations at petal fall to a few cells at harvest, with no detection of the pathogen after a 7-week cold storage. Migration of Ea153N into symptomless pear fruit from diseased branches was evaluated by enrichment assay and nested polymerase chain reaction of internal fruit core tissues; these assays failed to detect the pathogen in healthy fruit from diseased trees. At harvest, E. amylovora could not be detected on 5,599 of 5,600 fruit of d'Anjou pear sampled from commercial orchards in major production areas of the Pacific Northwest; one fruit yielded 32 CFU of the pathogen. Postharvest, mature pear fruit contaminated with Ea153N and subsequently wounded required a dose of >10,000 cells at the wound site to allow for persistence of the pathogen through a 7-week-cold storage. We conclude that epiphytic E. amylovora shows similar survival characteristics on both pear and apple fruit, this pathogen is not an endophyte within mature symptomless pear fruit, its presence is exceptionally rare on commercially produced fruit, and that epiphytic survival of E. amylovora through a postharvest chilling period is unlikely given the unrealistically high population size required for persistence.

15.
Am J Cardiol ; 98(3): 402-6, 2006 Aug 01.
Article in English | MEDLINE | ID: mdl-16860032

ABSTRACT

The accurate diagnosis of anomalous coronary arteries by invasive angiography is limited by the inability to define the anatomic course in relation to surrounding structures. Computed tomographic coronary angiography has recently emerged as a noninvasive method to visualize the coronary arteries. Multislice computed tomography with up to 64 detector arrays, along with 3-dimensional rendering, has further improved the temporal and spatial resolution of noninvasive coronary imaging. In this series of cases, the investigators describe their institution's experience with computed tomographic coronary angiography as a complement to invasive coronary angiography in determining the origin and course of different anomalous coronary arteries in 16 patients. With the aid of 3-dimensional volume rendering, 6 anomalous right coronary arteries, 4 anomalous left circumflex coronary arteries, 4 single coronary arteries, and 2 anomalous left main coronary arteries were all clearly defined with regard to their origin and course. It is proposed that computed tomographic coronary angiography is the diagnostic test of choice in the evaluation of such anomalies.


Subject(s)
Coronary Angiography/methods , Coronary Vessel Anomalies/diagnostic imaging , Tomography, X-Ray Computed , Adult , Aged , Diagnosis, Differential , Female , Follow-Up Studies , Humans , Male , Middle Aged , Reproducibility of Results , Retrospective Studies , Severity of Illness Index
16.
Phytopathology ; 95(9): 1072-80, 2005 Sep.
Article in English | MEDLINE | ID: mdl-18943305

ABSTRACT

ABSTRACT Efficiency of nonpathogenic Fusarium oxysporum Fo-B2 for the biological control of Fusarium wilt of tomato, caused by F. oxysporum f. sp. lycopersici CU1, was examined in different environments: a growth chamber with sterile soil-less medium, a greenhouse with fumigated or nonfumigated soil, and nonfumigated field plots. Inoculation of Fo-B2 onto tomato roots significantly reduced the severity of disease, but the efficiency of disease suppression decreased as the experimental environment became less controlled. Relationships between the recovery of Fo-B2 from hypocotyls and the disease severity indicated that the biocontrol agent was most effective when it colonized vascular tissues intensively. Moreover, the degree of Fo-B2 colonization was greatly reduced when the seedlings were grown in nonfumigated soil. Dose-response models (negative exponential, hyperbolic saturation, and logistic) were fit to observed data collected over a range of inoculum densities of the pathogen and the antagonist; the logistic model provided the best fit in all environments. The ratios of an 50% effective dose parameter for Fo-B2 to that of CU1 increased as the environment became less controlled, suggesting that environmentally related efficiency reduction impacted the antagonist more than the pathogen. The results suggest that indigenous soil microbes were a primary factor negatively influencing the efficiency of Fo-B2. Therefore, early establishment of the antagonist in a noncompetitive environment prior to outplanting could improve the efficacy of biological control.

17.
Am J Cardiol ; 93(10): 1318-9, 2004 May 15.
Article in English | MEDLINE | ID: mdl-15135717

ABSTRACT

Bivalirudin is being used more frequently as an anticoagulant in the cardiac catheterization laboratory. Newer devices, used to measure activated clotting time (ACT), have not been thoroughly tested for use with bivalirudin. One such device, the i-STAT ACT, measures the generation of activated thrombin to determine the level of anticoagulation. Our study demonstrated a high level of agreement between the i-STAT ACT and the Hemochron ACT in patients anticoagulated with bivalirudin. In addition, the i-STAT was shown to have an extremely high degree of reproducibility.


Subject(s)
Anticoagulants/blood , Blood Coagulation Tests/instrumentation , Hirudins/analogs & derivatives , Peptide Fragments/therapeutic use , Point-of-Care Systems/standards , Recombinant Proteins/therapeutic use , Cardiac Catheterization/methods , Humans , Reproducibility of Results
18.
Am J Cardiol ; 94(11): 1417-9, 2004 Dec 01.
Article in English | MEDLINE | ID: mdl-15566915

ABSTRACT

Three hundred nine patients were followed during their recovery area stay after percutaneous coronary intervention. Recovery area times for patients who received bivalirudin during percutaneous coronary intervention showed an average reduction in total recovery area length of stay of 36 minutes (p <0.0001) compared with patients who received heparin alone. This reduction was also seen when compared with the heparin + abciximab group (46-minute reduction, p = 0.0007), and the heparin + eptifibatide group (35-minute reduction, p = 0.0005). Patients who received bivalirudin took significantly less time for the activated clotting time (ACT) to normalize despite significantly higher average ACTs and significantly fewer subtherapeutic ACTs.


Subject(s)
Angioplasty, Balloon, Coronary , Antibodies, Monoclonal/therapeutic use , Anticoagulants/therapeutic use , Heparin/therapeutic use , Hirudins/analogs & derivatives , Immunoglobulin Fab Fragments/therapeutic use , Length of Stay , Peptide Fragments/therapeutic use , Peptides/therapeutic use , Recombinant Proteins/therapeutic use , Abciximab , Aged , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/pharmacology , Anticoagulants/administration & dosage , Anticoagulants/pharmacology , Antithrombins , Coronary Artery Disease/therapy , Drug Therapy, Combination , Eptifibatide , Female , Heparin/administration & dosage , Heparin/pharmacology , Hirudins/administration & dosage , Hirudins/pharmacology , Humans , Immunoglobulin Fab Fragments/administration & dosage , Immunoglobulin Fab Fragments/pharmacology , Male , Middle Aged , Peptide Fragments/administration & dosage , Peptide Fragments/pharmacology , Peptides/administration & dosage , Peptides/pharmacology , Recombinant Proteins/administration & dosage , Recombinant Proteins/pharmacology , Treatment Outcome
19.
Phytopathology ; 94(12): 1286-94, 2004 Dec.
Article in English | MEDLINE | ID: mdl-18943698

ABSTRACT

ABSTRACT The addition of 0.1 mM FeCl(3) to a defined culture medium induces the bacterial epiphyte Pseudomonas fluorescens strain A506 (A506) to produce an antibiotic toxic to the fire blight pathogen, Erwinia amylovora. Consequently, because A506 is registered and applied as a commercial product to suppress E. amylovora before floral infection of pear and apple, the relative availability of iron to A506 on surfaces of pear and apple flowers is of potential significance. An 'iron biosensor' construct of A506 was developed by transformation with an iron-regulated promoter (pvd) fused to a promoterless ice nucleation reporter gene (inaZ). This construct, A506 (pvd-inaZ), established high populations on pear and apple flowers, ranging from 10(4) to 10(6) CFU/flower. In seven trials on pear and apple trees, A506 (pvd-inaZ) expressed high ice nucleation activity (INA) on flowers, indicating limited iron bioavailability or a low-iron environment unlikely to induce antibiotic production by A506. A506 (pvd-inaZ) also colonized flowers when mixed with chemicals containing iron: FeSO(4) or the iron chelates ferric ethylenediaminedi-(o-hydroxyphenyl-acetic) acid (FeEDDHA) and ferric diethylenetriamine pentaacetate (FeDTPA). These compounds represent an array of commercial iron formulations applied to foliage to avert iron chlorosis. Treatment of flowers with a mixture of A506 (pvd-inaZ) and 3 mM FeEDDHA or FeDTPA significantly decreased INA compared with flowers treated with A506 (pvd-inaZ) in water. Lower concentrations (0.3 mM) of FeEDDHA, however, did not consistently suppress INA. These results indicate that apple and pear flowers represent an iron-limited environment to A506 and that treatment with 3 mM FeEDDHA is needed to increase significantly the level of iron available to this bacterium.

20.
Mol Plant Pathol ; 12(6): 580-94, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21722296

ABSTRACT

Xanthomonas hortorum pv. carotae (Xhc) is an economically important pathogen of carrots. Its ability to epiphytically colonize foliar surfaces and infect seeds can result in bacterial blight of carrots when grown in warm and humid regions. We used high-throughput sequencing to determine the genome sequence of isolate M081 of Xhc. The short reads were de novo assembled and the resulting contigs were ordered using a syntenic reference genome sequence from X. campestris pv. campestris ATCC 33913. The improved, high-quality draft genome sequence of Xhc M081 is the first for its species. Despite its distance from other sequenced xanthomonads, Xhc M081 still shared a large inventory of orthologous genes, including many clusters of virulence genes common to other foliar pathogenic species of Xanthomonas. We also mined the genome sequence and identified at least 21 candidate type III effector genes. Two were members of the avrBs2 and xopQ families that demonstrably elicit effector-triggered immunity. We showed that expression in planta of these two type III effectors from Xhc M081 was sufficient to elicit resistance gene-mediated hypersensitive responses in heterologous plants, indicating a possibility for resistance gene-mediated control of Xhc. Finally, we identified regions unique to the Xhc M081 genome sequence, and demonstrated their potential in the design of molecular diagnostics for this pathogen.


Subject(s)
Daucus carota/microbiology , Genome, Bacterial/genetics , Plant Diseases/microbiology , Sequence Analysis, DNA , Xanthomonas/genetics , Xanthomonas/pathogenicity , Base Sequence , Biomarkers , Genes, Bacterial/genetics , Genomics , Pathology, Molecular/methods , Phylogeny , Synteny/genetics , Nicotiana/microbiology , Virulence/genetics , Xanthomonas/classification , Xanthomonas/isolation & purification
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