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Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR 总被引:4,自引:1,他引:4
Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes . 相似文献
4.
Anthropogenic influences on potential fire spread in a pyrogenic ecosystem of Florida, USA 总被引:1,自引:1,他引:1
Fire has historically been an important ecological factor maintaining southeastern U.S. vegetation. Humans have altered natural
fire regimes by fragmenting fuels, introducing exotic species, and suppressing fires. Little is known about how these alterations
specifically affect spatial fire extent and pattern. We applied historic (1920 and 1943) and current (1990) GIS fuels maps
and the FARSITE fire spread model to quantify the differences between historic and current fire spread distributions. We held
all fire modeling variables (wind speed and direction, cloud cover, precipitation, humidity, air temperature, fuel moistures,
ignition source and location) constant with exception of the fuel models representing different time periods. Model simulations
suggest that fires during the early 1900's burned freely across the landscape, while current fires are much smaller, restricted
by anthropogenic influences. Fire extent declined linearly with patch density, and there was a quadratic relationship between
fire extent and percent landscape covered by anthropogenic features. We found that as little as 10 percent anthropogenic landcover
caused a 50 percent decline in fire extent. Most landscapes (conservation or non-conservation areas) are now influenced by
anthropogenic features which disrupt spatial fire behavior disproportionately to their actual size. These results suggest
that land managers using fire to restore or maintain natural ecosystem function in pyrogenic systems will have to compensate
for anthropogenic influences in their burn planning.
This revised version was published online in May 2005 with corrections to the Cover Date.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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C.?Silvar J.?M.?Duncan D.?E.?L.?Cooke N.?A.?Williams J.?Díaz F.?MerinoEmail author 《European journal of plant pathology / European Foundation for Plant Pathology》2005,112(1):43-52
A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils. 相似文献
6.
INVITED REVIEW—IMAGE REGISTRATION IN VETERINARY RADIATION ONCOLOGY: INDICATIONS,IMPLICATIONS, AND FUTURE ADVANCES 下载免费PDF全文
Yang Feng Jessica Lawrence Kun Cheng Dean Montgomery Lisa Forrest Duncan B. Mclaren Stephen McLaughlin David J. Argyle William H. Nailon 《Veterinary radiology & ultrasound》2016,57(2):113-123
The field of veterinary radiation therapy (RT) has gained substantial momentum in recent decades with significant advances in conformal treatment planning, image‐guided radiation therapy (IGRT), and intensity‐modulated (IMRT) techniques. At the root of these advancements lie improvements in tumor imaging, image alignment (registration), target volume delineation, and identification of critical structures. Image registration has been widely used to combine information from multimodality images such as computerized tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET) to improve the accuracy of radiation delivery and reliably identify tumor‐bearing areas. Many different techniques have been applied in image registration. This review provides an overview of medical image registration in RT and its applications in veterinary oncology. A summary of the most commonly used approaches in human and veterinary medicine is presented along with their current use in IGRT and adaptive radiation therapy (ART). It is important to realize that registration does not guarantee that target volumes, such as the gross tumor volume (GTV), are correctly identified on the image being registered, as limitations unique to registration algorithms exist. Research involving novel registration frameworks for automatic segmentation of tumor volumes is ongoing and comparative oncology programs offer a unique opportunity to test the efficacy of proposed algorithms. 相似文献
7.
Validity of goniometric joint measurements in cats 总被引:2,自引:0,他引:2
Jaeger GH Marcellin-Little DJ Depuy V Lascelles BD 《American journal of veterinary research》2007,68(8):822-826
OBJECTIVE: To compare and validate goniometric joint measurements obtained from nonsedated and sedated cats with measurements from radiographic evaluation. ANIMALS: 20 adult cats with no evidence of joint disease. PROCEDURES: Measurements of flexion and extension of the carpus, elbow, shoulder, tarsus, stifle, and hip joints and of carpal and tarsal joints during varus and valgus angulation were made by a single investigator before and after sedation of cats. Measurements were made by use of a goniometer with a masked dial. Joint angle measurements were compared between nonsedated and sedated cats and also with measurements from radiographs made while cats were sedated. Each series of measurements was repeated 4 times. To evaluate repeatability, Cronbach alpha values were calculated for repeated measure results of goniometric joint measurements of nonsedated and sedated cats. An intraclass correlation was calculated to determine reliability among the 3 measurement types (ie, measurements from nonsedated and sedated cats and on radiographic evaluation). RESULTS: Joint measurements did not differ significantly by measurement type, when comparing radiographic measurements with goniometric measurements in sedated and nonsedated cats. Cronbach alpha values were > 0.99 for goniometric joint measurements within individual nonsedated and sedated cats and also for comparison of mean measurements obtained from sedated cats versus nonsedated cats versus radiographs. An intraclass correlation of 0.999 revealed high reliability among measurement types. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that goniometric joint measurements in nonsedated and sedated cats are repeatable and valid. 相似文献
8.
OBJECTIVE: To determine historical, physical, and microbiologic findings in horses with limb cellulitis and identify factors associated with short- and long-term outcome. DESIGN: Retrospective case series. ANIMALS: 44 horses with limb cellulitis. PROCEDURES: Information obtained from medical records included use, history, affected limb, diagnostic procedures, treatment, and short-term outcome. Long-term follow-up information was obtained by means of a telephone survey. RESULTS: Twenty-four horses had primary cellulitis, and 18 of the 24 (75%) had a hind limb affected. Results of microbiologic culture were positive for all 20 horses with primary cellulitis from which specimens were obtained, with coagulase-positive Staphylococcus spp recovered from 12 of the 20. Short-term survival rate was 67% (16/24) for horses with primary cellulitis; 7 of the 9 horses available for long-term follow-up were being used for their intended use, and 4 had had a recurrence. Results of microbiologic culture were positive for 13 of the 16 (81%) horses with secondary cellulitis from which specimens were obtained. Short-term survival rate for horses with secondary cellulitis was 90% (18/20). Eleven of the 17 horses available for long-term follow-up were being used for their intended use; 2 had had a recurrence. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that cellulitis can be a life-threatening condition in horses. Horses that were febrile at admission or that developed laminitis were significantly less likely to survive. The prognosis for return to function was guarded, and recurrence was a potential concern. 相似文献
9.
To determine whether amino acid sequence variation exists in the Moraxella bovis (M. bovis) cytotoxin (MbxA) from geographically diverse M. bovis isolated in the United States, mbxA was amplified and sequenced. The MbxA deduced amino acid sequence from M. bovis originally isolated in California, Washington, North Carolina, and Georgia, as well as reference strains of M. bovis isolated at the National Animal Disease Laboratory, Ames, IA, USA, all encoded a nearly identical 927 amino acid protein. MbxA from two of the four California isolates (SFS 9a and SFS 100a) differed from all other isolates at two sites at which the polar amino acids glutamine (position 666) and asparagine (position 823) were replaced by ionized amino acids glutamic acid and aspartic acid, respectively. Rabbit antiserum to the expressed carboxy terminus (amino acids 590-927) of MbxA from M. bovis (Tifton I) neutralized the hemolytic activity of SFS 9a and SFS 100a. The M. bovis cytotoxin appears to be conserved amongst geographically diverse isolates of M. bovis from the USA. Antiserum against the carboxy terminus of MbxA common to the majority of isolates neutralized the hemolytic activity of two strains with a divergent MbxA deduced amino acid sequence. Vaccines against IBK that incorporate MbxA as antigen may offer protection against geographically diverse strains of M. bovis. 相似文献
10.
Kate L. Constantine Fernadis Makale Idah Mugambi Duncan Chacha Harrison Rware Alexander Muvea Vincent K. Kipngetich Justice Tambo Adewale Ogunmodede Djami Djeddour Corin F. Pratt Ivan Rwomushana Frances Williams 《Pest management science》2023,79(11):4343-4356